International Narcotics Research Conference

14 – 19 July 2013
Pullman Cairns International
Cairns, Australia

CONFERENCE HANDBOOK
www.dcconferences.com.au/inrc2013/www.dcconferences.com.au/inrc2013/

1

WELCOME
Welcome and thank you for participating in INRC 2013 in Cairns. This is the 44th meeting of the INRC. INRC has not met in Australia for\HDUV $GHODLGHLQ 7KLVFXUUHQWPHHWLQJLQ$XVWUDOLDDFNQRZOHGJHVWKHFRQWULEXWLRQV$XVWUDOLDQVFLHQWLVWVKDYHPDGHWRRXUÀHOGIn recognition of this, plenary lectures this year include presentations from two outstanding Australian scientists, Bernard Balleine andArthur Christopolous.
The highlight of our meeting as always is the Founder’s Lecture. This year the award goes to Dr. Graeme Henderson of the University
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Investigator Awardee, Dr. Louis Gendron of the Université de Sherbrooke, Canada.
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as well as the ever popular data-blitz. Please note that the symposium on “Endogenous Opioids” will be held in honor of one of INRC’s
founding fathers, Avram Goldstein, who passed away in 2012.
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inspires new ideas and collaborations.
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and we want to know your views.
Finally, I would like to thank Mark Connor and Macdonald Christie for putting together such a great program and for all the behind thescenes logistic work in getting this meeting off the ground. Of course, this meeting would not have been possible without our sponsors –the INRC thanks you.
Have a great meeting!

President, INRC

THANKS TO OUR SUPPORTERS

2

International Narcotics Research Conference 2013

CONTENTS
WELCOME

2

THANKS TO OUR SUPPORTERS

2

CONTENTS

3

COMMITTEES

4GENERAL INFORMATION

4SOCIAL PROGRAM

4PROGRAM

5

PROGRAM ABSTRACTS

7

POSTER LIST32

POSTER ABSTRACTS37

DELEGATE LIST

60

INDEX

63

NOTES

65

www.dcconferences.com.au/inrc2013/

3

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Committee
 Charles Chavkin, University of Washington, United States
 Ian Kitchen, University of Surrey, United Kingdom

Special thanks to the Local Organising
Committee
 Mark Connor, Co-Chair, Macquarie University, Australia
 Mac Christie, Co-Chair, University of Sydney, Australia
 Andrew Lawrence, Florey Institute/University of Melbourne,
Australia
 Ernie Jennings, James Cook University at Cairns, Australia
 Andrew Somogyi, University of Adelaide, Australia

 Christoph Stein, Freie Universität Berlin, Germany Susie Ingram, Oregon Health and Science University, UnitedStates John Traynor, University of Michigan Medical School, UnitedStates Louis Gendron, Université de Sherbrooke, Canada Lan Ma, Fudan University Pharmacology Research Center,
China Alfreda Stadlin, Chungbuk National University, South Korea

 Minoru Narita, Hoshi University School of Pharmacy andPharmaceutical Science, Japan

GENERAL INFORMATIONSOCIAL PROGRAM

Venue

Welcome Reception

Pullman Cairns International

Date:

17 Abbott Street

Venue: Daintree Pool Deck, Pullman Cairns International

CAIRNS QLD 4870

Time:

T: +61 7 4031 1300

Soak up the delights of tropical Cairns whilst enjoying a refreshing
beverage or two on the Daintree Pool Deck. Take time out tocatch up with friends and colleagues.

F: +61 7 4031 1801
W: www.pullmancairnsinternational.com.au

Registration Inclusions
Delegate registration includes:

Sunday 14 July 2013
6.00pm – 8.00pm

The Welcome Reception is complimentary for all conferencedelegates. Additional tickets can be purchased for $60 per personat the registration desk.

 $WWHQGDQFHWRDOOVFLHQWLÀFVHVVLRQV

Conference Data Blitz

 6DWFKHO LQFOXGLQJFRQIHUHQFHPDWHULDOV

Date: :HOFRPH5HFHSWLRQ 6XQGD\HYHQLQJ

Venue: Mossman Ballroom, Ground Floor Pullman Cairns
International

 0RUQLQJWHDOXQFKDQGDIWHUQRRQWHD 0RQGD\²7KXUVGD\DQG
morning tea on Friday)

Monday 15 July 2013

Time:

6.30pm – 8.30pm

 &RQIHUHQFHKDQGERRN

3ULFH

Complimentary

Registration Desk Opening Hours

The INRC Conference Data Blitz offers students and postdocs the
chance to introduce results in a casual session. All students andpostdocs are welcome to present their data.

Sunday 14 July: 4.00pm – 6.00pm
Monday 15 July: 7.30am – 5.30pm
Tuesday 16 July: 8.00am – 6.00pm
Wednesday 17 July: 8.00am – 12.30pm
Thursday 18 July: 8.00am – 5.00pm
Friday 19 July: 8.00am – 12.30pm

The presentation is a starting point for informal discussions withlight refreshments to follow.

Conference Dinner
Date:Thursday 18 July 2013

Venue: Thornton Peak, Pullman Cairns International
Time:

7.00pm – 10.30pm

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3ULFH

$80 per person

Opening hours-RLQ\RXU,15&FROOHDJXHVIRUDQHYHQLQJRIJUHDWIRRGÀQHZLQH
DQGH[FHOOHQWFRQYHUVDWLRQXQGHUWKHVWDUVRIDEDOP\WURSLFDO
North Queensland evening.

Sunday 14 July: 4.00pm – 5.30pm
Monday 15 July: 7.00am – 2.00pm
Tuesday 16 July: 7.00am – 2.00pm

Please visit the registration desk to purchase tickets to theconference dinner.

Wednesday 17 July: 7.00am – 11.00am

4International Narcotics Research Conference 2013

Welcome Reception: 6pm – 8pm Daintree Pool Deck, Pullman Cairns International

Monday, 15 July 2013
TIME

PRESENTATION

8.00 – 8.30am

Arrival Tea and Coffee

SPEAKER

Welcome8.30 – 9.45am

9.45am
9.45 – 10.10am
10.10 – 10.35am
10.35 – 11.15am
11.15 – 11.40am

Proudly sponsored by the Ian Potter Foundation

3OHQDU\ Founders Lecture: Opioid research – turning fullcircleSymposium 1: Opioid heteromers
Chair: Lakshmi Devi

Graeme Henderson

Proudly sponsored by the British Pharmacological Society and the British Journal of
Pharmacology

Co-localization of Mu and Delta Opioid Receptors in the Nervous System Using Double
Fluorescent Knock-in Mice
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tolerance
Morning Tea
Symposium 1 Continued: Opioid heteromers
A µOR-įOR Heteromer-biased ligand with antinociceptive activity

Dominique Massotte
Xu Zhang

Wakako Fujita

11.40amSymposium 2: Human Aspects of Opioid Use

11.40 – 12.05pm

Heroin dependence in Asian populations

Alfreda Stadlin

12.05 – 1.00pm

Andrew Somogyi

1.25 – 1.50pm

Lunch
Symposium 2 Continued
Opioid-induced hyperalgesia: does the myth need busting?
Patterns of prescription drug use in Australia

1.50 – 2.15pm

Afternoon Tea

2.15 – 3.15pm

Hot Topics 23.30 – 5.30pm

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1.00 – 1.25pm

Paul Haber
Kate Dolan, Alisa Knapman,
6KX/XQJ<DQJ1D\OD&KDLMDOH

6.30pm – 8.30pm Data Blitz Mossman Ballroom, Ground Floor Pullman Cairns International

Tuesday, 16 July 2013
TIME

PRESENTATION

8.00 – 8.30am
8.30amArrival Tea and Coffee
Symposium 3: MOR regulation
Chair: Macdonald Christie

8.30 – 8.55am

Differentiation of opioid drug effects by hierarchical multi-site phosphorylation

Stefan Schulz

8.55 – 9.20am

0RGXODWLRQRI025ELQGLQJDIÀQLW\E\GHVHQVLWL]LQJDJRQLVWVWill Birdsong

Proudly sponsored by the British Pharmacological Society and the British Journal of
Pharmacology

9.45 – 10.15am

Acute injury establishes constitutive µ-opioid receptor activity leading to long-term
endogenous analgesia and dependence
Morning Tea

10.15 – 11.00am

Hot Topics 311.00amSymposium 4: Opioids in pain states
Chair: Susan Ingram

11.00 – 11.25am

Amygdala plasticity as a mediator of diverse persistent pain conditions

9.20 – 9.45am

SPEAKER

Bradley Taylor$UVDODQ<RXVXI6HNVLULArttamangkul, Janet Lowe

Robert Gerreau IV

11.50 – 12.15pm

Spinal cord AMPA receptors containing the GluA4 subunit mediate morphine-induced
hyperalgesia
Opioid reward in pain states

12.15 – 1.15pm

Lunch

1.15 – 2.00pm

Hot Topics 42.00pmSymposium 5: Opioids In non-neuronal cells

2.00 – 2.25pm

Role of glia in opioid-mediated analgesia

Sandra Comer

2.25 – 2.50pm

Microglia-mediated disruption of neuronal Cl – homeostasis gates morphine hyperalgesia

Tuan Trang

2.50 – 3.15pm

Morning Tea

3.15 – 3.40pm

Opioid Actions in Non-Neuronal Cells – The Heart

3.40 – 4.25pm

Hot Topics 54.30 – 6.00pm

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11.25 – 11.50am

Jose Moron-Concepcion
Minoru Narita
Karen Tonsfeldt, BenjaminLau, Kelly StandiferJohn Headrick
Marie-Odile Parat, Mei
Bigliard, Naoko Kuzumaki

PROGRAM

Sunday, 14 July 2013

PROGRAM

Wednesday, 17 July 2013
TIME

PRESENTATION

8.00 – 8.30am

Arrival Tea and Coffee
3OHQDU\8.30 – 9.30am

9.30amSPEAKER

Proudly sponsored by the International Society for Neurochemistry

Allostery and bias at G protein-coupled receptors. Implications for
drug discovery
Symposium 6: Bias/allostery
Chair: Mark Connor

Arthur Christopoulos

Proudly sponsored by the International Society for Neurochemistry

9.30 – 9.55am

Measurement and mechanism of ligand bias at MOPr

10.20 – 11.00am

Ligand and cell-dependent determinants of cellular responses to delta opioid receptor
ligands
Morning Tea

11.00 – 12.15pm

Hot Topics 6

12.15pm

Lunch9.55 – 10.20am

Eammon Kelly
Graciela Pineyro
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John Traynor, James Zadina,
Erin Bobeck

Thursday, 18 July 2013
TIME

PRESENTATIONSPEAKER

8.00 – 8.30am

Arrival Tea and Coffee
3OHQDU\Forebrain mechanisms of opioid action: Reward and stimulus control ofchoice and decision-making
Symposium 7: Endogenous Opioids Chair: Larry Toll

Bernard Balleine

8.30am – 9.30am

Proudly sponsored by The American Society for Pharmacology and Experimental
Therapeutics in memory of Avram Goldstein

9.30 – 9.55am
9.55 – 10.20am
10.20 – 10.50am
10.50 – 11.20am

Endogenous Opioids in the Amygdala
Opioid peptides induce long-term depression at glutamatergic synapses in the dorsal
striatum
Morning Tea

Elena Bagley
Brady Atwood

Hot Topics 7
Symposium 8: INRC Young Investigator Symposium
Chair: John Traynor%ULDQ&R[$\D0DWVXL

11.25 – 12.00pm

Roles and regulation of the delta opioid receptor in pain

Louis Gendron

12.00 – 12.25pm

ȕ-arrestin 1 control of delta receptor and ORL signaling

Wendy Walin

12.25 – 2.00pm

Lunch and Poster Presentations

2.00 – 2.25pm'25PDWXUDWLRQDQGWUDIÀFNLQJ

2.25 – 3.10pm

Hot Topics 8

3.10 – 3.45pm

Afternoon Tea

3.45 – 5.00pm

INRC Business Meeting

7.00 – 10.30pm

Conference Dinner

11.20 – 11.25am

Ulla Petaja-Repo
Erica Levitt, Karim Nagi,
Bronwyn Kivell

Friday, 19 July 2013
TIME

PRESENTATION

8.00 – 8.30am

8.55 – 9.20am

Arrival Tea and Coffee
Symposium 9: Hypocretin/Orexin and Addiction
Chair: Ingrid NylanderMorphine-induced plasticity of Ventral Tegmental Area dopamine
QHXURQVLVJDWHGE\+\SRFUHWLQ2UH[LQUHFHSWRUVLJQDOLQJThe hypocretin-dynorphin system in reward and drug-seeking

9.20 – 9.45am

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9.45 – 10.15am

Hot Topics 910.15 – 10.45am

Morning Tea
Symposium 10: Mechanisms of opioid synergy
Chair:Thomas Tzschentke

8.30am8.30 – 8.55am

10.45am

SPEAKER

Stephanie Borgland
John Muchamp
Selena Bartlett
Lih-Chu Chiou
Andrew Lawrence

Proudly sponsored by Grunenthal

10.45 – 11.10am

A subset of spinal analgesic synergy requires activation of protein kinase C-epsilon*HRUJH:LOFR[

11.10 – 11.35am

Cannabinoid/opioid interactions: mechanisms for improving pain relief

Adie Wilson-Poe

11.35 – 12.00pm([SORLWLQJEHWZHHQRSLRLGGLIIHUHQFHVWRHQKDQFHDQDOJHVLDDQGLPSURYHWROHUDELOLW\

Maree Smith

12.00 – 12.15pm

Hot Topics 10

Mahsa Sadeghi

PROGRAM
ABSTRACTS
3/(1$5<
23,2,'5(6($5&+²7851,1*)8//&,5&/(Henderson, G.
School of Physiology & Pharmacology, University of Bristol, BS8 1TD, UK

The aim of this lecture will be to describe how the use of electrophysiological recording techniques have contributed to ourunderstanding of the mechanisms underlying the profound effects of opioid drugs on the central nervous system.
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crystal structures even, and use more sophisticated tools but the same questions still remain to be answered.

3/(1$5<$//267(5<$1'%,$6('$*21,60$7*3&5V5(/(9$1&(72129(/'58*',6&29(5<Christopoulos, A.
Monash Institute of Pharmaceutical Sciences & Dept. of Pharmacology, Monash University,VIC, 3052, Australia

G protein-coupled receptors (GPCRs) constitute the largest family of drug targets. In the new millennium, two important paradigms have
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(or functional selectivity), which posits that different ligands engender unique receptor conformations such that only a subset of pathways
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implications with respect to screening for, or interpreting structure-function studies of, novel GPCR-targeting small molecules. The
development of novel analytical approaches for quantifying allosteric and biased ligand effects on GPCRs, as well as new breakthroughs in
GPCR structural and computational biology, promise to facilitate new approaches to enriching structure-activity relations and structurebased drug design at this therapeutically important receptor family.

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23,2,'352&(66(6,15(:$5'$1''(&,6,210$.,1*
Balleine, B.
Brain & Mind Research Institute, University of Sydney, NSW, Australia

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however, much less well understood, whether the focus is at a behavioral, neural systems or cellular level. Although often construed interms of hedonics, reward signals mainly regulate the evaluative processes through which the goals of goal-directed actions are acquired
on which decision-making processes operate. As a consequence, it would appear likely that opioid-related processes play a central role in
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(ii) the way reward and reward prediction modulate decision-making at both a behavioural and a cellular level.

www.dcconferences.com.au/inrc2013/7

PROGRAM ABSTRACTS

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&2/2&$/,=$7,212)08$1''(/7$23,2,'
5(&(37256,17+(1(592866<67(086,1*
'28%/()/825(6&(17.12&.,10,&(
Massotte, D.
Dept Neurobiology and Translational Medicine, IGBMC, Illkirch, France:HKDYHJHQHUDWHGGRXEOHNQRFNLQDQLPDOVH[SUHVVLQJGHOWDRSLRLGUHFHSWRUVLQIXVLRQZLWKWKHJUHHQÁXRUHVFHQWSURWHLQDQG
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used these animals to map the two receptors in the brain, spinal
cord and dorsal root ganglia and create a virtual atlas of their
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FRH[SUHVVLQJWKHWZRUHFHSWRUVDUHLQGHHGSUHVHQWLQDQXPEHU
of brain structures. This suggests potential physiological relevanceand prompts to investigate the reality of in vivo functional mu-delta
heteromers to assess them as new potential therapeutic targets.

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Institute of Neuroscience and State Key Laboratory of Neuroscience,
Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences,
Shanghai, 200031, China

7KHH[FLWDWRU\QHXURWUDQVPLVVLRQRIQRFLFHSWLYHDIIHUHQWWHUPLQDOVin the dorsal horn of spinal cord can be modulated by the
inhibitory opioid peptides released from the local neurons andH[RJHQRXVO\DSSOLHGRSLRLGDQDOJHVLFVVXFKDVPRUSKLQHWKURXJK
activating the presynaptic į- and µ-opioid receptors (DORs and
025V UHVSHFWLYHO\:HÀQGWKDW025VDQG'25VDUHFR
H[SUHVVHGLQDFRQVLGHUDEOHSRSXODWLRQRIQRFLFHSWLYHQHXURQVLQthe dorsal root ganglion (DRG). However, these types of opioid
receptors have distinct subcellular distributions. MORs aremainly localized on the cell surface, while DORs mostly distributein the cytoplasm and are often localized in the membrane of
neuropeptide-containing large dense-core vesicles. The vesicular
localization of DORs allows activity-dependent membrane
insertion of DORs and interaction with MORs in the plasma
membrane. The function of MORs could be negatively regulated by'25V$FWLYDWLRQRI'25VLQWKH'25025FRPSOH[UHVXOWVLQ
co-internalization and degradation of DORs and MORs. Morphineanalgesic tolerance could be reduced by treatment with DOR
antagonists or disruption of the DOR/MOR interaction in the
nociceptive afferent terminals.

8$25į25+(7(520(5%,$6('/,*$1':,7+
$17,12&,&(37,9($&7,9,7<*Fujita, W.1, Gomes, I.1, Gupta, A.1+RGGHU32, Roberts, E.3,
and Devi, L.A.1
1
Department of Pharmacology and Systems Therapeutics, Icahn School of
Medicine at Mount Sinai, New York, NY 10029, 2 Department of Molecular
Therapeutics,The Scripps Research Institute, Jupiter, FL 33458 and, 3 Scripps
Research Institute Molecular Screening Center, La Jolla, CA 92037.

Emerging studies suggest that dimerization of G protein-coupled
receptors (GPCRs) including opioid receptors is necessary for
UHFHSWRUPDWXUDWLRQVLJQDOLQJDQGWUDIÀFNLQJ+HWHURPHUL]DWLRQ
between opioid receptor (OR) types is thought to affect thepharmacological effects of opiates. We have previously shownthat µOR and įOR form heteromers whose presence canbe detected in brain regions involved in pain processing usingµOR-įOR heteromer-selective antibodies. We also reported
that chronic morphine administration leads to an upregulation
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distinct from that of µOR or įOR alone. These results suggestthat the µOR-įOR heteromer could serve as a novel therapeutictarget for the treatment of chronic pain. Here we report theLGHQWLÀFDWLRQRIFRPSRXQGVVSHFLÀFDOO\WDUJHWLQJ25įORheteromers by high-throughput screening of a library of small
molecules using a ȕDUUHVWLQUHFUXLWPHQWDVVD\:HÀQGWKDWRI
WKHaFRPSRXQGVWHVWHGH[KLELWUHFHSWRUDFWLYDWLRQRQO\LQFHOOVH[SUHVVLQJ25į25EXWQRWLQFHOOVH[SUHVVLQJ25RU
į25DORQH6LQFHRIFRPSRXQGVH[KLELWHGOHVVWKDQ
structural similarity to known opioids, these were repurchased
and reanalyzed using in vitro and in vivo assays. Among the top 3
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įOR-biased ligand. Furthermore, intrathecal administration
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can be blocked by µOR-įOR heteromer selective antibodies.
Interestingly, while the antinociceptive activity of systemically
DGPLQLVWHUHG&<0LVVLPLODUWRWKDWRIPRUSKLQHLWH[KLELWV
lesser antinociceptive tolerance compared to morphine. TheseUHVXOWVVXJJHVWWKDW&<0D25įOR-biased ligand,could serve as a scaffold for the development of a novel type of(heteromer-biased) drug that is as potent as the conventional
clinical opioids with reduced side-effects. Supported by NIH grants
NIH grants DA008863 and DA019521 (L.A.D.) and by the NIH
Molecular Library Screening Center Network Grant MH084512(P.H. and E.R.).

International Narcotics Research Conference 2013

PROGRAM ABSTRACTS

6<0326,80

,038/6,9,7<3(5621$/,7<75$,76$1'*(1(32/<0253+,606$662&,$7(':,7++(52,1
'(3(1'(1&(Dissabandara, L.O.1,2, Loxton, N.J.3, Ho, A.MC.2'RGG354,
Daglish, M.2, *Stadlin, A.5
6FKRRORI0HGLFLQH*ULIÀWK8QLYHUVLW\*ROG&RDVW2 School of Medicine,
Psychology and 4 Chemistry & Molecular Biosciences,The University of
4XHHQVODQG+HUVWRQ$XVWUDOLD5 Chungbuk National University School of
Medicine, Cheongju, South Korea
1
3

Impulsivity is a risk factor associated with all facets of addiction
from initial predisposition, through the maintenance phase and asa predictor of relapse risk. Genes encoding molecular pathways of
the reward system are implicated in both addiction and impulsivity.
$FRPSOH[LQWHUSOD\RIWKHVHJHQHVLVOLNHO\WRFRQWULEXWHWRWKH
risk of addiction, both directly and indirectly, mediated via theintermediate phenotype of impulsivity. We assessed the impulsivity
personality traits and nineteen candidate polymorphisms of the
GABA, opioid and dopamine pathways of the reward system inan ethnically distinct and homogenous population sample of 157
Sinhalese, male dependent heroin users and 155 age- and ethnicityPDWFKHGH[WUHPHFRQWUROVZLWKQROLIHWLPHGUXJXVHDQGQHJOLJLEOH
nicotine or alcohol consumption, using direct, indirect and epistatic
association analyses. Results showed that the use of a twofactor model of impulsivity provided additional, novel important
information about substance-use behaviours. The diagnosis ofdependence was associated with both higher rash-impulsivenessand reward sensitivity, while heroin dependence-associated subphenotypes such as high-risk behaviour that included escalating
heroin consumption, injecting heroin use, hazardous drinking, low
WUHDWPHQWVHHNLQJDQGULVN\VH[XDOEHKDYLRXUZHUHDVVRFLDWHG
only with high rash-impulsiveness. An early onset of drug use wasassociated with reward sensitivity. By applying different analytic
strategies based on direct, indirect and epistatic associations topreviously reported and/or functionally relevant candidate variants
we can enhance the biological plausibility of their associations
and increase the chances of revealing subtle polygenetic effects.
GABA receptor SNPs (GABRG2-rs3219151 and GABRG2-rs211013)
showed both individual and haplotype association with heroin
dependence. Two opioid mu-receptor SNPs (OPRM1-rs1799971
and OPRM1-rs563649) also showed individual association withheroin dependence. The same GABA SNPs and DRD2-rs1079597
were associated with impulsivity-related traits, which mediated theeffect of these polymorphisms on heroin dependence. Further, we
IRXQGDVWDWLVWLFDOO\VLJQLÀFDQWDQGELRORJLFDOO\UHOHYDQWHSLVWDWLF
interaction between GABRG2 and OPRM1 for risk of heroin
GHSHQGHQFH7KHVHÀQGLQJVSRUWUD\QRYHODQGSODXVLEOHSRWHQWLDO
mechanisms of genetic predisposition to heroin dependence.

23,2,',1'8&('+<3(5$/*(6,$'2(67+(0<7+1(('%867,1*"
Somogyi, A.
Discipline of Pharmacology, School of Medical Sciences University of
Adelaide, Adelaide 5005 Australia
Opioid-induced hyperalgesia (OIH) is a phenomenon whereby
RSLRLGVVHHPWRLQGXFHDSDUDGR[LFDOLQFUHDVHLQSDLQVHQVLWLYLW\(and more diffuse pain) and is seen after chronic opioid dosing
in dependence and pain treatments and sometimes in the acuteperioperative setting. The somewhat “mythical” nature of OIH is incontrast to opioid-induced tolerance (OIT) whereby an increase in

www.dcconferences.com.au/inrc2013/

dose is needed to achieve the same analgesic effect. Using the coldpressor model, ‘tolerance’ time is markedly reduced in methadone
and buprenorphine maintained subjects and in chronic pain
patients after all opioids tested. IOH and IOT can be detected also
after prolonged intraoperative dosing with high potency opioids
IHQWDQ\ODQGUHPLIHQWDQLO )RU2,+LWVFOLQLFDOH[LVWHQFHLVGHEDWHG
in terms of patient group (acute versus cancer versus noncancerpain), detection methods, and relationship to opioid dosing routeand regimen.
As not all people develop IOH (or IOT), the question of agenetic risk factor has been raised but without resolution or
research. Nevertheless for both IOH and IOT, the landscapeof the molecular mechanism has mainly involved the neuronalsystem (NMDA receptor, glutamate, dynorphin, mu G-protein).
However, the role of the CNS innate immune system and especially
SURLQÁDPPDWRU\F\WRNLQHVPD\EHLPSRUWDQWDVPDQ\DQLPDODQG
human studies and case reports have shown that ketamine (theVRFDOOHG10'$DQWDJRQLVW FDQDWWHQXDWHWRDFHUWDLQH[WHQW
both IOH and IOT during acute and chronic opioid administration.
.HWDPLQHKDVDQWLSURLQÁDPPDWRU\HIIHFWVYLDVWHUHRVHOHFWLYH
DQGSRVVLEO\VWHUHRVSHFLÀFHIIHFWVRQ7/50'DQGUHGXFWLRQLQ
LPS-induced IL-6 secretion. The busting of the myth of OIH and its
mechanism(s) may herald new approaches to how opioids are usedin the clinical setting.

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36<&+2$&7,9(0(',&$7,216,1$8675$/,$+DEHU31, Blanch, B.2
Drug Health Services, Royal Prince Alfred Hospital, University of Sydney,
NSW Australia 2 School of Pharmacy, University of Sydney, NSW Australia
1

The use of pharmaceutical opioid drugs in the community is
FRQWURYHUVLDOZLWKHYLGHQFHRIERWKVLJQLÀFDQWXQGHUXWLOL]DWLRQDQG
RYHUXWLOL]DWLRQ7KHUHLVVRPHHYLGHQFHWRVXSSRUWHIÀFDF\IRU
chronic non-malignant pain, but community and individual harms
are increasingly reported. There is an increasing range of otherpsycho-active medications that may contribute to therapeuticEHQHÀWDQGWRRYHUGRVHULVN7KHDLPRIWKLVSUHVHQWDWLRQLVWR
review availability of opioid and other psycho-active prescription
SDWWHUQVWKURXJKWKHVXEVLGL]HG3KDUPDFHXWLFDO%HQHÀWV6FKHPH
in Australia from 1992 to the present. Methods: Interrogation of
WKH3KDUPDFHXWLFDO%HQHÀWV6FKHPHRQOLQHVWDWLVWLFVGDWDIURP
1992 onwards. Item numbers for all available preparations were
LGHQWLÀHGDQGIUHTXHQF\RIGLVSHQVLQJZDVFROOHFWHGDQGFROODWHG
Results: The number of medication classes, and the number ofpreparations within each class increased during this period, in thecase of opioids from 11 to 70 preparations. The total number of
PBS opioid prescriptions increased from 6.0 million in 1992 to 18
million in 2012, a three-fold increase. There were dramatic and
FRQWLQXLQJLQFUHDVHVLQSUHVFULSWLRQRIR[\FRGRQHEXSUHQRUSKLQH
and fentanyl but little change in use of morphine and methadone.
There were also dramatic increases in dispensing of olanzapine,quetiapine and risperidone. Conclusion: There is increasing use of
DOOFODVVHVRIPHGLFDWLRQVH[DPLQHG7KHUHLVDSUHVVLQJQHHGWR
implement strategies to improve the quality usage of opioids andother psycho-active medications, individually and in combination.

9

PROGRAM ABSTRACTS+27723,&6

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HCV SEROINCIDENCE
Dolan, K.1, Salimi, S.2, Nassirimanesh, B.3, Mohsenifar, S.2,
Allsop, D.1, Mokri, A.2
Program of International Research and Training, National Drug and Alcohol
5HVHDUFK&HQWUH$XVWUDOLD2 Iranian National Center for Addiction Studies,
7HKUDQ8QLYHUVLW\RI0HGLFDO6FLHQFHV,UDQ3 Persepolis Centre,Tehran, Iran1

Background: In general, information about women who use drugs
comes from studies performed in the West. Whether women incountries such as Iran are likely to enter drug treatment or how
they will respond is not known.
3XUSRVH7RH[DPLQHWKHVKRUWWHUPLPSDFWRIPHWKDGRQH
maintenance treatment (MMT) on drug use, dependence, social
IXQFWLRQLQJFULPHDQGKXPDQLPPXQRGHÀFLHQF\YLUXV +,9 DQG
hepatitis C virus (HCV) risk behavior and seroincidence in female
drug users in Iran.
Methods: Women were eligible for inclusion in the study ifthey were assessed as dependent on opiates according to the,QWHUQDWLRQDO6WDWLVWLFDO&ODVVLÀFDWLRQRI'LVHDVHVDQG5HODWHG
Health Problems, tenth revision (ICD-10). The sample comprised
78 female heroin or opium users who attended the Persepolis
women’s drug treatment clinic in Tehran between 2007 and 2008.
Participants were followed up in 2009/2010. Heroin and theuse of other drugs, social functioning, involvement in crime, andinvolvement in HIV and HCV risk behavior were measured by
self-report. The prevalence and incidence of HIV and HCV were
measured by serology and self-report.
Findings: Of the 78 women recruited, 40 were followed up, andWKLVRFFXUUHGDSSUR[LPDWHO\PRQWKVODWHU2QHLQIRXUZRPHQ
reported a history of drug injection. At follow-up there were
VLJQLÀFDQWUHGXFWLRQVLQVHOIUHSRUWHGKHURLQXVHRQ,&'
dependence scores. Subjects with more severe drug dependence
DWEDVHOLQHZHUHVLJQLÀFDQWO\PRUHOLNHO\WREHFULPLQDOO\DFWLYH
than less severely dependent subjects. Baseline prevalence for+,9DQG+&9ZDVDQGUHVSHFWLYHO\$WIROORZXSQRRQH
had acquired HIV infection, but one participant had acquired HCV,
giving an incidence rate of 7.1 per 100 person-years.
Conclusion: 7KLVUHVHDUFKSURYLGHVWKHÀUVWHYLGHQFHWKDW,UDQLDQ
female drug users can enter MMT and respond well. Within a few
months of entering MMT, improvements occurred in heroin use,levels of dependence, social functioning, and HIV risk behavior.
While the incidence of blood-borne viral infections was low, there
was a serious risk of HIV transmission among this cohort and
DOVRWRSDUWLFLSDQWV·QHHGOHDQGVH[XDOFRQWDFWV,QDFRXQWU\ZLWKhigh levels of drug use, the high levels of HCV among female drug
users require more women to enter drug treatment if an HIV
epidemic is to be avoided. Many participants had a chronic drug
SUREOHPDQGKDGKDGOLWWOHRUQRSUHYLRXVH[SRVXUHWR0077KH
LQWURGXFWLRQRUH[SDQVLRQRIZRPHQRQO\GUXJWUHDWPHQWVHUYLFHV
is urgently needed in order to engage more women in treatment.

10

7+(())(&72)23,2,'5(&(3725
32/<0253+,606215(&(3725)81&7,21*Knapman, A., Connor, M.
The Australian School of Advanced Medicine, Macquarie University, Sydney,
NSW, Australia

7KHUHLVVLJQLÀFDQWYDULDWLRQLQLQGLYLGXDOUHVSRQVHWRRSLRLG
drugs, one cause of which is likely to be polymorphisms on the
opioid receptors themselves. The µ-opioid receptor (MOR) is
the primary site of action for most analgesic opioids. Previous
VWXGLHVKDYHLGHQWLÀHGDQXPEHURIQDWXUDOO\RFFXUULQJVLQJOH
nucleotide polymorphisms (SNPs) in the coding region of the
MOR. The A118G SNP variant, present in various populations
DWDOOHOLFIUHTXHQFLHVUDQJLQJIURP²KDVEHHQDVVRFLDWHG
with differences in the requirement for post-operative opioid
analgesics, and for drug dependence, although these results are not
FRQVLVWHQW,QYLWUR025$*KDVEHHQUHSRUWHGWRH[KLELWDQ
DOWHUHGVLJQDOOLQJSURÀOHFRPSDUHGZLWK025LQFOXGLQJGLIIHUHQWȕ-endorphin potency and alterations in N-type Ca2+ channel
LQKLELWLRQKRZHYHUWKHVHÀQGLQJVDUHDOVRQRWFRQVLVWHQW)HZVWXGLHVKDYHH[DPLQHGWKHHIIHFWRI025SRO\PRUSKLVPVRQ
receptor function, or potential ligand functional selectivity. We
DVVHVVHGWKHUHODWLYHSRWHQF\DQGHIÀFDF\RIDUDQJHRIRSLRLG
ligands in Chinese hamster ovary (CHO) cells stably transfected
ZLWKKXPDQZLOGW\SH025DQGÀYHQDWXUDOO\RFFXUULQJ025
YDULDQWVLQFOXGLQJ$*5HFHSWRUH[SUHVVLRQOHYHOVZHUHVLPLODUIRUDOOPXWDQWVH[DPLQHG025PHGLDWHGDGHQ\O\OF\FODVH
$& LQKLELWLRQZDVPHDVXUHGXVLQJDQRYHOÁXRUHVFHQFHEDVHG
membrane potential assay. Treatment of CHO cells with the AC
activator forskolin (FSK) hyperpolarized CHO-MOR cells with aS(&RIWRDPD[LPXPRIIURPEDVHOLQH7KH
hyperpolarization induced by FSK (300nM) was inhibited in a dosedependent manner by the addition of a range of MOR agonists,
including DAMGO, morphine, ȕ-endorphin and buprenorphine (n ,Q025$*FHOOVWKHPD[LPDOEXSUHQRUSKLQHLQKLELWLRQ
RI$&ZDVUHGXFHGWR FRPSDUHGZLWKLQ025
WT, P < 0.05), without a change in potency, while responses to
PRUSKLQHDQG%HQGZHUHXQDIIHFWHG&HOOVH[SUHVVLQJ025&7VKRZHGVLJQLÀFDQWO\UHGXFHG(PD[DQGS(&IRUPRUSKLQH
but not DAMGO. By contrast, MOR-C253A cells showed
VLJQLÀFDQWO\LQFUHDVHG(PD[IRUPRVWOLJDQGVZLWKQRFKDQJHLQ
SRWHQF\&+2025&7ZDVFRQÀUPHGDVQRQIXQFWLRQDO7KH
differences in inhibition of AC observed between MOR variants
suggest that MOR SNPs may contribute to individual variability in
WKHUHVSRQVHWRRSLRLGDQDOJHVLFVDQGZHDUHH[SORULQJWKLVDWDrange of signalling pathways.

International Narcotics Research Conference 2013

PROGRAM ABSTRACTS+27723,&6

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TAIWAN

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02'8/$7,212)7+(/2&86&2(58/(86
125(3,1(3+5,1(6<67(0$1',1&5($6(6THE SALIENCE OF REWARD.

<DQJ6KX/XQJ*Chaijale, N., Curtis A.L., Snyder, K., Bhatnagar,
S.,Valentino, R.J.

Crime Research Center, National Chung Cheng University, Chia-Yi. Taiwan

The effectiveness of drug addicts ‘ two different models (i.e.,
institutional treatment-compulsory treatment in drug abuser
treatment center and deferred prosecution for drug addicts’
treatment in the community) for schedule I offenders (majorly
Heroine) has been empirically tested and evaluated by the author 6SHFLÀFDOO\WKHVWXG\VDPSOHGUXJDGGLFWVIURPdrug abuser treatment center, and 362 clients under deferred
prosecution order, for the evaluation of their effectiveness in terms
RIIDPLO\DWWDFKPHQWIULHQGDWWDFKPHQWDGGLFWLRQVHYHULW\LQGH[and Short Form-36(Physical, mental and social assessment) for theyear of 2012.This study found that the two groups in normal family
attachment, normal friend attachment and quality of life were
VLJQLÀFDQWO\LPSURYHGDIWHUWUHDWPHQWEXWDOVRVLJQLÀFDQWO\UHGXFH
DGGLFWLRQDVPRVWVWXGLHVH[SHFWHG%XWWKHWZRJURXSVLQDQ
attempt to reduce both drug addicts’ family attachment, and drug
addicts’ friend attachment , to improve get rid of the motivations
RIGUXJXVHDQGTXDOLW\RIKHDOWKDUHQRWVWDWLVWLFDOO\VLJQLÀFDQWNevertheless, both of institutional treatment and deferred
prosecution treatment are generally effective.
Despite these, the effectiveness of such comparison has not\HWEHHQHPSLULFDOO\YHULÀHGIRUVFKHGXOH,,RIIHQGHUV PDMRUO\$PSKHWDPLQH LQ7DLZDQ7KHFXUUHQWVWXG\DWWHPSWVWRÀOOVXFKvoid, and provided further evidence. This study conducts a seriesÀHOGZRUNVXUYH\VWRDQDO\]HWKHRXWFRPHVDQGFRVWEHQHÀW
HIIHFWLYHQHVVRIWZRPRGHORIWUDGLWLRQDOGHWHQWLRQGHWR[LÀFDWLRQ
group(schedule II drug addicts of Taipei Detention Center andTaipei Women’s Detention Center) and deferred prosecution
and addiction treatment group(schedule II drug addicts of Taipei'LVWULFW3URVHFXWRUV2IÀFHDQG7DLSHL&LW\+RVSLWDO6RQJGHEUDQFK). After analyzing the data, the policy implications of two judicial
models can be derived.

www.dcconferences.com.au/inrc2013/Childrens Hospital of Philadelphia, Philadelphia, PA. USA.

The locus coeruleus-norepinephrine (LC-NE) system is amajor stress response system through which stress affects
arousal and cognitive function. The LC is co-regulated by stressneuromediators and endogenous opioids. Because social stress(SS) is a common stressor for humans, this study characterized
WKHHQGXULQJLPSDFWRIUHSHDWHG66RQ/&DFWLYLW\6SHFLÀFDOO\ZH
determined whether SS alters the LC-NE system two (Day 7) andten days (Day 15) after the last SS and evaluated whether thesechanges in LC-NE system translate to changes in cognitive function.
Rats were implanted with a multiwire bundle into the LC andH[SRVHGWRÀYHVHVVLRQVRIWKHUHVLGHQWLQWUXGHUPRGHORIVRFLDO
defeat. On Day 7, LC discharge rate was decreased in stressed rats
compared to controls (Day 1: 2.21±0.1Hz, Day 7: 1.35±0.1Hz forstress rats and Day 1: 2.23±0.1Hz, Day 7: 2.20±0.1Hz, for control
rats; n<40). By Day 15, LC rates were comparable between groups.
6\VWHPLFDGPLQLVWUDWLRQRIWKHRSLDWHDQWDJRQLVWQDOR[RQHUREXVWO\increased LC rate selectively in stressed rats when administered
RQ'D\ +]SUHQDOR[RQH+]SRVWQDOR[RQH 
RU'D\ +]SUHQDOR[RQH+]SRVWQDOR[RQH 
This cellular evidence of opiate withdrawal was accompanied bybehavioral signs of mild opiate withdrawal. To evaluate whether
66LQGXFHGFKDQJHVLQ/&DFWLYLW\DIIHFWFRJQLWLYHÁH[LELOLW\UDWV
were tested in an attention set shifting task (AST) and LC activity
was recorded. Stress rats performed better in intradimensional
set shifting (IDS): 36.7±4 and 23.8±4 trials to criterion for control
and stress rats, respectively (p<0.05). Recordings during IDS
showed that LC neurons of stressed rats were reward, but notdecision responsive. During reversal learning although overall
performance was comparable between groups, stressed rats madeless perseverative errors. LC neurons of stressed rats selectively
showed decision- and reward-related activation. LC activity also
increased for stressed rats only during incorrect trials between
decision and recognition of the absence of reward. These results
suggest that SS engages endogenous opioid modulation of LC
activity and induces a state of opiate dependence. Additionally, SS
strengthens the relationship between LC activity and goal-directedbehavior. Together these effects of SS on the LC system canincrease vulnerability to opiate abuse by promoting the positive
and negative reinforcing effects. Supported by T32-NS007413,
58077 LSDRP, MH40008, DA09082.

11

PROGRAM ABSTRACTS

6<0326,80

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())(&76%<+,(5$5&+,&$/08/7,6,7(
3+263+25</$7,21
Just, S., Illing, S.,7UHVWHU=HGOLW]0Lau,E.K., Kotowski,S.J.,
Miess, E., Mann,A., Doll, C.,Trinidad, J.C., Burlingame, A.L.,
YRQ=DVWURZ0Schulz, S.
Institute of Pharmacology and Toxicology (S.J., S.I., E.M., A.M., C.D., and S.S.),
Jena University Hospital, Friedrich Schiller University Jena, Drackendorfer
Straße 1, D-07747 Jena, Germany.
Departments of Psychiatry (M.T.-Z., E.K.L., S.J.K., and M.v.Z.), Pharmaceutical
Chemistry (J.C.T., and A.L.B.), Chemistry (A.L.B.), and Cellular and Molecular
Pharmacology (M.v.Z.), University of California, San Francisco, CA 94158,
USA.

Differences in the ability of opioid drugs to promote regulated
endocytosis of µ-opioid receptors are related to their tendency to
produce drug tolerance and dependence. Here we show that drugVSHFLÀFGLIIHUHQFHVLQUHFHSWRULQWHUQDOL]DWLRQDUHGHWHUPLQHGE\D
FRQVHUYHGUHVLGXHVHTXHQFHLQWKHUHFHSWRU·VFDUER[\OWHUPLQDO
cytoplasmic tail. Diverse opioids induce receptor phosphorylation
at S375, present in the middle of this sequence, but opioids differ
markedly in their ability to drive higher-order phosphorylation onÁDQNLQJUHVLGXHV 77DQG7 0XOWLSKRVSKRU\ODWLRQLV
required for the endocytosis-promoting activity of this sequenceand occurs both sequentially and hierarchically, with S375
representing the initiating site. Higher-order phosphorylation
LQYROYLQJ77DQG7VSHFLÀFDOO\UHTXLUHV*5.
isoforms, and the same sequence controls opioid receptor
internalization in neurons. These results reveal a biochemicalmechanism differentiating the endocytic activity of opioid drugs.

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3+263+25</$7,21,1&5($6(6$*21,67
$)),1,7<$77+(0823,2,'5(&(3725'2(6,7$))(&7'(6(16,7,=$7,21"
Birdsong, W.T.,1 Arttamangkul, S.,1YRQ=DVWURZ02,
Williams, J.1
Vollum Institute, Oregon Health & Science University, Portland, OR, USA.
University of California San Francisco, San Francisco, CA, USA

1
2

3URORQJHGH[SRVXUHWRKLJKHIÀFDF\DJRQLVWVUHVXOWVLQUHFHSWRU
phosphorylation at the mu opioid receptor (MOR) anddesensitization of downstream signaling. It is unknown what effects
GHVHQVLWL]LQJDJRQLVWH[SRVXUHKDVRQDJRQLVWUHFHSWRULQWHUDFWLRQV8VLQJFRQIRFDOLPDJLQJDQGDÁXRUHVFHQWDJRQLVW GHUPRUSKLQ
$OH[D ZHIRXQGWKDWSURORQJHGH[SRVXUHWRGHVHQVLWL]LQJDJRQLVWVLQFUHDVHGWKHDIÀQLW\RI025IRUGHUPRUSKLQ$OH[D7KLVLQFUHDVHLQDIÀQLW\ZDVORQJODVWLQJLQGHSHQGHQWRI
downstream signaling and abolished by mutation of putative
phosphorylation sites. Thus, conditions that are known to cause
receptor desensitization also result in receptor phosphorylation
FDXVLQJDQLQFUHDVHGDJRQLVWDIÀQLW\7KHUROHRIUHFHSWRU
phosphorylation in opioid signaling will be discussed.
Supported by DA08163, DA026617 (JTW) and DA007262-18(WTB).

$&87(,1-85<(67$%/,6+(6&2167,787,9(µ23,2,'5(&(3725$&7,9,7</($',1*72
/21*7(50(1'2*(1286$1$/*(6,$$1'
'(3(1'(1&(Taylor, B.
Injury causes acute pain sensation that resolves with healing;
however, the transition to a persistent pathological pain stateremains an important clinical problem. Endogenous activation ofµ-opioid receptors (MORs) provides relief from acute pain, but
WKHH[LVWHQFHRIORQJWHUPRSLRLGLQKLELWLRQRISDWKRORJLFDOSDLQKDVQRWEHHQVXIÀFLHQWO\H[SORUHG+HUHZHVKRZWKDWDFXWHWLVVXH
LQÁDPPDWLRQSURGXFHVDJRQLVWLQGHSHQGHQW025VLJQDOLQJLQWKH
dorsal horn, which tonically represses spinal nociceptive signaling
for months. Disruption of this constitutive activity reinstated signsof pain and neuronal sensitization, and precipitated cellular, somaticand aversive signs of opioid withdrawal. This required N-methyl-Daspartate receptor activation of calcium-sensitive adenylyl cyclase
type 1. Thus, we have discovered a novel mechanism of long-lasting
opioid analgesia that regulates the transition from acute to chronic
pain while, in parallel, generates opioid dependence. The prevalenceof chronic pain syndromes may result from a failure in constitutive
signaling of spinal MOR.

12International Narcotics Research Conference 2013

PROGRAM ABSTRACTS+27723,&6

7+(52/(2)',))(5(173+263+25</$7,21
6,7(6,15$3,''(6(16,7,=$7,212)7+(23,2,'5(&(3725<RXVXI$1, Miess, E.2, Schulz, S.2, Christie, M.J.1
Discipline of Pharmacology, University of Sydney, NSW, Australia, 2Instituteof Pharmacology and Toxicology, Jena University Hospital, Friedrich Schiller
University Jena, Jena Germany

1

Tolerance and addiction to opioids are serious clinical andsocial problems that result in part from the loss of µ-opioid
receptor (MOR) function when activated by different opioids.
3KRVSKRU\ODWLRQRIVSHFLÀFUHVLGXHVLQWKH&WHUPLQDOGRPDLQRI
MOR is thought to be a key step in desensitization, endocytosis
and loss of MOR function. Sequential phosphorylation of S375
DORQJZLWKWKHÁDQNLQJUHVLGXHV77DQG7DUHUHTXLUHG
for endocytosis (Just et al., Mol Pharmacol (2013) 83:633-639)
but their role in rapid desensitization of MOR is unknown. The
DLPRIWKLVVWXG\ZDVWRGHWHUPLQHWKHLQÁXHQFHRI&WHUPLQDO
phosphorylation sites on rapid desensitization of MOR. Wild typemMOR, 3S/T-A (S363A, S370A, S375A), 6 S/T-A (S363A, S370A,
S375A, T376A, T379A, T383A) and 11S/T-A (T354A, S355A, S356A,
T357A, S363A, S370A, S375A, T376A, T379A, T383A, T394A)
ZHUHH[SUHVVHGVWDEO\LQ$W7FHOOV8VLQJSHUIRUDWHGSDWFKFODPSUHFRUGLQJZHH[DPLQHGWKHHIIHFWVRI025DFWLYDWLRQdesensitization and re-sensitization using activation of GIRKFKDQQHOVE\DVXEPD[LPDOFRQFHQWUDWLRQRIPHWHQNHSKDOLQ nM) to measure receptor activity and somatostatin (100 nM)
coupling to native SSTR2 (or sst2 according to IUPHAR) to
determine heterologous desensitization. MOR desensitization and
UHVHQVLWL]DWLRQSURGXFHGE\PLQH[SRVXUHWRPHWHQNHSKDOLQ
(10 µM) at 37oC did not differ from wild type for 3S/T-A or
6S/T-A but desensitization was abolished in the 11S/T-A mutant.
Desensitization, when detected, was largely homologous. Because
3S/T-A suppresses, and 6S/T-A abolishes MOR endocytosis, theseÀQGLQJVVXJJHVWWKDWKRPRORJRXVGHVHQVLWL]DWLRQFDQRFFXU
independently of the phosphorylation and arrestin-dependent
mechanisms that drive endocytosis. However, C-terminal
phosphorylation sites are necessary for desensitization because
mutation of all C-terminal sites (11S/T-A) abolishes desensitization.

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)520'(6(16,7,=$7,21,1/2&86
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Arttamangkul, S.1, Schulz, S.2 and Williams, J.T. 1
1Vollum Institute, Oregon Health & Science University, Portland, Oregon, USA
2Pharmacology, Jena University Hospital, Jena, Germany

The actin cytoskeleton acts to support cellular structure, but it
also plays a role in GPCR signaling and regulation. The network
RIDFWLQÀODPHQWVEHQHDWKSODVPDPHPEUDQHKDVEHHQVKRZQWRFRQÀQHPXRSLRLGUHFHSWRUV 025 LQWRDFRPSDUWPHQWDQG
restrains global lateral diffusion properties. The present study
used latrunculin B (Lat B) to disrupt actin polymerization in
HEK293 cells or mouse locus coeruleus neurons (LC). The effectof Lat B on lateral diffusion of Flag-tagged MOR (Flag-MOR) in+(.FHOOVZDVVWXGLHGE\OLQHVFDQQLQJÁXRUHVFHQFHFRUUHODWLRQ
spectroscopy. In Lat B treated cells, the diffusion rate of Flag-MOR
was faster than control. The effect of Lat B on [Met5]enkephalin(ME)-induced receptor internalization was studied in brain slices of
ORFXVFRHUXOHXVQHXURQVIURPWUDQVJHQLFPLFHH[SUHVVLQJ)ODJ

www.dcconferences.com.au/inrc2013/

MOR. In the Lat B treated cells the amount of internalized FlagMOR and the endosome size was increased compared to control
following treatment with ME (30 µM, 10 min). The desensitization
measured by the decline in the hyperpolarization induced by ME(30 µM) was not altered but there was only a partial recovery
from desensitization following the wash. The impaired recovery
from desensitization correlated with a reduction in the recycling
RIUHFHSWRUVEDFNWRWKHSODVPDPHPEUDQH([SHULPHQWVXVLQJ/&slices obtained from mutant S375A knock-in mice showed that thedesensitization and the recovery were not altered by Lat B. This
study suggests that the actin cytoskeleton plays a key role in the
UHFRYHU\IURPGHVHQVLWL]DWLRQE\UHJXODWLQJWKHUHFHSWRUWUDIÀFNLQJThe study is supported by NIH grant DA08163.

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0823,2,'5(&(37256'(3(1'6217+(,5&(//8/$5/2&$/,=$7,21
*Lowe, J.D., and %DLOH\&3
Dept. of Pharmacy & Pharmacology, University of Bath, Bath, United Kingdom

A primary site for opioid-induced euphoria is activation of muopioid receptors (MOPrs) in the ventral tegmental area (VTA). The
causes and mechanisms of tolerance to opioid-induced euphoria
are poorly understood, but one hypothesis for tolerance in generalis agonist-induced MOPr desensitization. In the VTA, MOPrs arelocated on GABAergic interneurons both somatodendritically(ie. on cell bodies and dendrites) and on nerve terminals thatinnervate dopaminergic neurons. Using whole-cell patch-clamp
electrophysiological methods, we have investigated agonist-induced
desensitization of both populations of MOPrs in mouse VTA slices
DQGIRXQGSURIRXQGGLIIHUHQFHVLQWKHLUGHVHQVLWL]DWLRQSURÀOH
depending on their subcellular localization. In common with manyother neuronal cell-types, agonist-induced MOPr desensitizationis readily seen at VTA cell bodies, where the DAMGO-induced
UHVSRQVHGHFOLQHGE\DSSUR[LPDWHO\GXULQJDPLQXWH
application. In contrast, neither DAMGO, Met-Enkephalin, normorphine were able to induce rapid MOPr desensitization at
nerve terminals even in the absence of a receptor reserve.
Because activation of PKC has been shown to enhance morphine
mediated desensitization in other brain regions (Bailey et al.
2004), we treated slices with the phorbol ester PMA. Althoughmorphine still did not appear to induce desensitization during a
10 minute application, PMA treatment dramatically reduced thepeak morphine response compared to untreated slices suggesting
desensitization may have occurred during the slow onset ofmorphine action (> 5 minutes). PMA did not affect the peak
response to DAMGO, nor promote DAMGO desensitization overthe 10-minute application. Therefore, PKC selectively enabledmorphine-induced desensitization of nerve terminal MOPrs.
$OWKRXJKWKHKLJKHIÀFDF\023UDJRQLVWV'$0*2DQG0HW
Enkephalin were unable to promote rapid desensitization of nerve
terminal receptors, preliminary data suggest that treating slices for
7-10 hours with Met-Enkephalin reduced nerve terminal MOPr
function in most cells. Thus, MOPrs located at nerve terminals
GHVHQVLWL]HYHU\VORZO\WRKLJKHIÀFDF\DJRQLVWVEXWGRGHVHQVLWL]Hto morphine if PKC is activated. Overall, these results demonstrateprofound differences in the mechanism of MOPr desensitization
based both on their localization within the neuron and on which
agonist is used to activate them. (Funded by MRC)
Bailey CP et al. (2004) Mol. Pharm., 66, 1592-98.

13

PROGRAM ABSTRACTS

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',9(56(3(56,67(173$,1&21',7,216
Gerreau, R. IV
The comorbidity of multiple chronic pain conditions and thepresence of enhanced sensitivity at sites far removed from thesite of tissue injury are consistent with an anatomical locus of
SDLQDPSOLÀFDWLRQZLWKLQWKHFHQWUDOQHUYRXVV\VWHP,QWKLV
presentation, I will describe studies performed in our lab utilizing
anatomy, behavioral pharmacology, and optogenetic approaches todemonstrate a role for mGluR5-dependent signaling in the central
QXFOHXVRIWKHDP\JGDODLQWKHFRQWH[WRISHUVLVWHQWSDLQ7KHGDWD
support the hypothesis that plasticity in central amygdala leads
to the generation of widespread pain in response to anatomically
restricted injury.

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81'(5/<,1*0253+,1(,1'8&('3$,1
Cabanero, D.1, Carlton, S.2, *Concepcion, J.M.1
Dept Anesthesiology, Columbia University, New York2
Dept Neuroscience, University of Texas Medical Branch, Galveston,Texas

1,QGLYLGXDOVZKRGLVFRQWLQXHRSLRLGXVHDIWHUUHSHDWHGH[SRVXUH
develop an abstinence syndrome in which one of the core
symptoms is an increase in pain sensitivity. In patients treatedwith opioids, this hypersensitivity can increase the probability ofabuse. It is thought that the molecular mechanisms underlying
opioid-induced hypersensitivity are regulated by plasticity events
that converge at the dorsal horn of the spinal cord. We haverecently reported that repeated morphine administration triggers
an insertion of GluA2-lacking (Ca2+-permeable) AMPA receptors$03$5 LQWKHKLSSRFDPSXV7KLVÀQGLQJWRJHWKHUZLWKWKH
reported involvement of AMPAR in the mechanisms underlyingLQÁDPPDWRU\SDLQOHGXVWRK\SRWKHVL]HDUROHIRUVSLQDO$03$5
in opioid-induced pain behavior. Mice treated with escalating dosesof morphine showed hypersensitivity to mechanical stimulation.
Intrathecal administration of the Ca2+-permeable AMPAR selective
blocker, naspm, disrupted morphine-induced mechanical sensitivity.
$QDO\VLVRIWKHH[SUHVVLRQDQGSKRVSKRU\ODWLRQOHYHOVRI$03$5
subunits (GluA1/2/3/4) in homogenates and in postsynaptic density
fractions from spinal cord dorsal horns showed an increase in*OX$H[SUHVVLRQDQGSKRVSKRU\ODWLRQLQWKHSRVWV\QDSWLFGHQVLW\after morphine. Co-immunoprecipitation analyses suggested anincrease in GluA4 homomers (Ca2+-permeable AMPAR) andimmunohistochemical staining localized the increase in GluA4OHYHOVLQODPLQDH,,,9,QDGGLWLRQWKHH[FLWDWRU\SRVWV\QDSWLF
currents (EPSCs) recorded in laminae III-V showed enhanced
sensitivity to Ca2+-permeable AMPAR blockers in morphine
treated mice. Furthermore, current-voltage relationships of$03$5PHGLDWHG(36&VVKRZHGWKDWUHFWLÀFDWLRQLQGH[ DQ
indicator of Ca2+-permeable AMPAR contribution) is increased inmorphine-treated, but not saline-treated mice. Interestingly, theseeffects could be reversed by infusion of GluA4 antibody throughpatch pipette. Therefore, we propose that discontinuation ofmorphine treatment induces pain sensitivity through the synaptic
insertion of GluA4-containing Ca2+-permeable AMPAR at spinal
cord dorsal horn neurons. Overall, this study highlights spinal
GluA4-containing AMPAR as new targets to prevent the pain
sensitivity that develops following the discontinuation of opioid
treatment.

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TRANSMISSION*Narita, M.,<DPDVKLWD$
Dept. Pharmacol., Hoshi Univ. Sch. Pharm. Pharmaceut. Sci., 142-8501, Japan

Although morphine and other mu-opioid agonists are the main
analgesics for severe pain, these compounds have potential forabuse and/or addiction. However, clinical studies show that whenmu-agonist analgesics are appropriately used to control pain, actual
DEXVHRUDGGLFWLRQGRHVQRWXVXDOO\RFFXU&OLQLFDOH[SHULHQFH
and animal studies support the contention that opioid euphoria
DQGUHZDUGDUHDWWHQXDWHGE\SDLQ,QÁDPPDWRU\QRFLFHSWLRQ
and neuropathic pain resulted in the suppression of activatedmesolimbic dopaminergic transmission evoked by mu-opioids.
These results indicate that chronic nociceptive stimuli may induce
the functional down-regulation of the rewarding network, leading
to the depression and a reduction in abuse potential of mu-opioid
analgesics. We also found that these phenomena were eliminated
LQPLFHWKDWODFNHGWKHEHWDHQGRUSKLQJHQH1H[WZHXVHG
optogenetic techniques to selectively inhibit VTA-NAc pathway.
To evaluate the effect of inhibiting the VTA-NAc pathway on
WKHH[SUHVVLRQRIDQDOJHVLDSURGXFHGE\PRUSKLQHZHLQMHFWHG
AAV-halorhodopsin into the VTA of TH-Cre mice in vivo. Phasic
VWLPXODWLRQGHFUHDVHG97$GRSDPLQHQHXURQDOH[FLWDELOLW\DQG
inhibited antinociception produced by morphine during optical
VWLPXODWLRQ7KHVHÀQGLQJVSURYLGHHYLGHQFHIRUWKHFUXFLDOUROH
of the activated endogenous mu-opioidergic system in negatively
controlling the abuse potential of opioids by chronic nociceptive
stimuli. In this symposium, we will discuss our current data andreview the molecular mechanisms of pain-induced suppression of
opioid dependence.

Supported by NIH grant R01DA027460 to JMC.
14International Narcotics Research Conference 2013

PROGRAM ABSTRACTS+27723,&6

CHRONIC INFLAMMATION ALTERS*$%$6<1$37,&$&7,9,7<$1'08
23,2,'5(&(3725,1+,%,7,21,17+(3(5,$48('8&7$/*5$<*Tonsfeldt, K.J., Suchland, K.L., Li. M & Ingram, S.L.
Dept. of Neurological Surgery, Oregon Health & Science University, Portland,
OR&KURQLFLQÁDPPDWRU\SDLQFDQLQGXFHQHXURDGDSWDWLRQVZLWKLQthe central nervous system. Opioid inhibition of GABAergic
synaptic activity in the periaqueductal gray (PAG) activates adescending antinociception pathway that reduces nociceptive
neurotransmission in the spinal cord. Little is known aboutchanges in synaptic transmission in the PAG following chronicLQÁDPPDWLRQ0HWHQNHSKDOLQ 0( LQKLELWLRQRI3$**$%$HUJLF
V\QDSWLFWUDQVPLVVLRQZDVH[DPLQHGLQPDOHDQGIHPDOHUDWV
pretreated with Complete Freud’s Adjuvant (CFA) or saline.
Recordings from brain slices containing the PAG were madeGD\VIROORZLQJLQMHFWLRQV,QIHPDOHVWKH&)$LQÁDPHGSDZ
PHDVXUHGPPDQGZDVVLJQLÀFDQWO\ODUJHUWKDQWKH
uninjected hindpaw (3.7 ± 0.2 mm, t(5) = 6.293, p < 0.05) withno observed change in saline injected hindpaws. Basal GABAmIPSC frequency was increased in CFA (2.2 ± 0.5 Hz, n = 12 cells)
compared to saline pretreated rats (1.1 ± 0.2 Hz, n = 8 cells; MannWhitney U = 26, p < 0.05). Inhibition of mIPSC frequency by ME(300 nM and 10 µM) was enhanced in CFA-treated rats (26 ± 5
DQGQ FHOOV FRPSDUHGWRVDOLQH DQG
n = 9 cells). These results suggest that CFA induces an increase
in GABA release and increases ME inhibition of GABA mIPSC
frequency.
Interestingly, different results were observed in male rats
pretreated with CFA. Although there was a similar increase in paw
diameter with CFA injections (6.0 ± 0.2 mm, N = 4) compared tothe control paws (3.6 ± 0.2 mm, t(3) = 14.10, p < 0.05), the basal
mIPSC frequency in male rats trended toward decreased frequency
in CFA pretreated rats (0.9 ± 0.2 Hz, n = 8 cells) compared tosaline (2.8 ± 1.3 Hz, n = 3 cells). In contrast to the female rats,
there was no change in ME (300 nM and 10 µM) inhibition of
P,36&IUHTXHQF\LQ&)$SUHWUHDWHGUDWV DQGQ
FHOOV FRPSDUHGWRVDOLQH DQGQ FHOOV 
Thus, GABA inhibition in the PAG is modulated differently by CFALQGXFHGLQÁDPPDWLRQEHWZHHQPDOHDQGIHPDOHUDWV6XSSRUWHGE\DA027625 (SLI).

www.dcconferences.com.au/inrc2013/

23,2,'',6,1+,%,7,212)7+('(6&(1',1*$1$/*(6,&3$7+:$<,17+(3(5,$48('8&7$/*5(<
Lau, B.K., & Vaughan, C.W.
Kolling Institute of Medical Research, Royal North Shore Hospital, University
of Sydney

3XUSRVHThe midbrain periaqueductal grey (PAG) is a majorsite of analgesic action by opioids. These agents have long been
hypothesized to produce analgesia via an indirect process of‘disinhibition’ in descending analgesic systems. Of particular interestis the descending pathway in the PAG, which projects throughthe rostral ventromedial medulla (RVM) to modulate nociceptive
transmission at the spinal cord dorsal horn. While many studies
suggest that µ-opioids activate the PAG-RVM descending pathway
via a disinhibitory cellular mechanism, there is no evidence to datedirectly demonstrating suppression of the inhibitory inputs onto
PAG output neurons projecting to the RVM. Thus, there is still a
ODFNRIGHÀQLWLYHVXSSRUWIRUGLVLQKLELWLRQRIWKHSULQFLSDOQHXURQV
involved in pain modulation. The present study aimed to address
WKLVLVVXHE\H[DPLQLQJWKHFHOOXODUDFWLRQVRIRSLRLGVVSHFLÀFDOO\RQLGHQWLÀHG3$*590RXWSXWQHXURQV
Methods: PAG output neurons projecting to the RVM wereretrogradely labelled in Sprague-Dawley rat pups (17-20 days old).
Electrophysiological whole-cell patch clamp recordings were then
FRQGXFWHGIURPWKHVHLGHQWLÀHG3$*RXWSXWQHXURQVUHFRUGLQJHYRNHGLQKLELWRU\DQGH[FLWDWRU\SRVWV\QDSWLFFXUUHQWV ,36&V
EPSCs). The frequency and amplitude of the individual quantal
HYHQWVXQGHUO\LQJWKHHYRNHG,36&V(36&VZHUHH[DPLQHGXVLQJVWURQWLXP P0 LQWKHH[WHUQDOUHFRUGLQJVROXWLRQResults: The µ- and Ɉ-opioid agonists, DAMGO and U69593
produced a reduction of both evoked IPSCs and EPSCs in labelled
PAG output neurons, which was associated with paired pulse
facilitation. In contrast, the Ɂ-opioid agonist, deltorphin-II had
QRVLJQLÀFDQWHIIHFWVRQV\QDSWLFWUDQVPLVVLRQ7KHUHVSRQVHRI
'$0*2DQG8RQ(36&VZDVVLJQLÀFDQWO\OHVVFRPSDUHG
to their effect on IPSCs. DAMGO was also found to reduce thefrequency of Sr2+-mediated evoked miniature IPSCs in labelled
output neurons.
Conclusions: Hence, µ- and Ɉ-opioid agonists act via apresynaptic mechanism to suppress both inhibitory GABAergic and
H[FLWDWRU\JOXWDPDWHUJLFV\QDSWLFWUDQVPLVVLRQRQWR3$*RXWSXW
neurons. We have previously shown that only a small proportion of
output neurons in the ventrolateral PAG respond directly to opioid
DJRQLVWV7RJHWKHUWKHVHÀQGLQJVVXJJHVWWKDWRSLRLGVDFWLQGLUHFWO\
via a disinhibitory action to activate the PAG-RVM descending
pathway. This is consistent with the opioid disinhibition model of
DQDOJHVLD)XUWKHUPRUHWKHREVHUYHGVXSSUHVVLRQRIH[FLWDWRU\V\QDSWLFWUDQVPLVVLRQOLNHO\VXJJHVWVFRPSOH[HIIHFWVE\RSLRLGVLQ
this analgesic system.

15

PROGRAM ABSTRACTS+27723,&6

3+<6,2/2*,&$/$1'3$7+23+<6,2/2*,&$/
5(*8/$7,212),/(;35(66,21%<
12&,&(37,1253+$1,1)4 12)4 3(37,'(5(&(3725
Donica, C.L.2=KDQJ<16LPSVRQ'XUDQG&'1, *Sherry,
D.M.2, 3, Gallucci, R.M.1, 2, Awwad, H.O.1, 2 and Standifer,
K.M.1, 2, 3
Department Pharmaceutical Sciences, College of Pharmacy, 2 Oklahoma
Center for Neuroscience and 3 Department Cell Biology, College of Medicine,
University of Oklahoma HSC, OKC, OK, USA
1

The neuropeptide, N/OFQ, has immunomodulatory actions in
DGGLWLRQWRPRGXODWLQJQRFLFHSWLRQDQ[LHW\DQGPDQ\RWKHUUHVSRQVHV([SUHVVLRQRI,QWHUOHXNLQ ,/ DF\WRNLQHOLQNHGWRLQÁDPPDWLRQDQGSDLQLVPRGXODWHGE\12)412)4SHSWLGH(NOP) receptor signaling at the mRNA and protein levels. To
better understand how N/OFQ modulates IL-6, IL-6 mRNA levels
in U937 human monocyte cells were assessed by quantitative
PCR (qPCR). Time course studies revealed that N/OFQ initially
increased IL-6 mRNA at 15 min, but decreased IL-6 mRNA over
WKHQH[WKU3UHWUHDWPHQWZLWKD123UHFHSWRUDQWDJRQLVW
blocked the N/OFQ-induced increase in IL-6 mRNA at 15 min,with no effect on the subsequent reduction in IL-6 at 4 hr. N/
OFQ-induced up-regulation of IL-6 mRNA also was blocked byinhibitors of NFɈB, p38 and Akt signaling, but not by inhibitorsof PKC or ERK MAP kinase; suggesting potential mechanisms bywhich N/OFQ increases IL-6. We previously reported that Single
Prolonged Stress (SPS), a PTSD model, produced hyperalgesia andallodynia, and increased N/OFQ levels in serum, CSF and brain.
7RH[SORUHSRWHQWLDOUHODWLRQVKLSVEHWZHHQ12)4F\WRNLQHV
and PTSD-related pain, we assessed IL-6 mRNA and protein
levels in the spinal cord dorsal horn of male Sprague-Dawley rats
H[SRVHGWR636DQGPDWFKLQJFRQWUROVE\T3&5PXOWLSOH[F\WRNLQH
analysis and immunolabeling. SPS increased IL-6 mRNA in spinal
cord and brain between days 14-28 days, but IL-6 levels in sera
were unchanged. Preliminary studies indicate that treatment of
SPS rats with a NOP receptor antagonist from day 7-21 blocked
IL-6 protein up-regulation in spinal cord at day 21 and reversed
hyperalgesia and allodynia, suggesting that N/OFQ mediates IL-6
up-regulation during chronic stress and that this up-regulation may
contribute to allodynia and hyperalgesia in the PTSD model.
Animal protocol was approved by IACUC and the US Army
Medical Research and Materiel Command Animal Care and Use
5HYLHZ2IÀFHFRPSOLHGZLWKWKH$QLPDO:HOIDUH$FWDQGDGKHUHG
to the principles described in the Guide for Care and Use ofLaboratory Animals.

16

International Narcotics Research Conference 2013

PROGRAM ABSTRACTS

6<0326,80

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0(',$7('5(63216(6,1+80$15(6($5&+
92/817((560,&52*/,$0(',$7('',65837,212)
1(8521$/&/+20(267$6,6*$7(60253+,1(+<3(5$/*(6,$*Comer, S.D., Jones, J.D., Mogali, S., Manubay, J., Sullivan,
M.A., &RRSHU='Vosburg, S.K., Metz,V., Johnson, K.W.

Trang,T.

Columbia University and the New York State Psychiatric Institute, New York,NY USA

Several recent preclinical studies have suggested that glia may play
an important role in opioid-mediated effects, including analgesia,
UHVSLUDWLRQDQGUHZDUG6SHFLÀFDOO\LQKLELWLRQRIJOLDODFWLYDWLRQLQFUHDVHVWKHSRWHQF\DQGHIÀFDF\RIRSLRLGLQGXFHGDQDOJHVLDDQG
simultaneously reduces opioid-induced reward and respiratory
depression. Glia also modulate opioid-induced tolerance anddependence in preclinical models. However, the effects of inhibitionof glial activation on opioid-induced responses in humans have
not yet been investigated. Two glial attenuators (ibudilast andminocycline) were tested in a laboratory setting in combination
ZLWKRSLRLGVXQGHUDYDULHW\RIH[SHULPHQWDOFRQGLWLRQV,EXGLODVWa non-selective phosphodiesterase-4 and -10 and macrophagemigration inhibitory factor (MIF) inhibitor, is used clinically in
Asia to treat asthma and post-stroke dizziness, and minocycline,
a tetracycline antibiotic, is used to treat acne and other bacterial
infections. The effects of ibudilast on opioid withdrawal symptoms
in morphine-dependent heroin abusers were investigated, as was
WKHDELOLW\RILEXGLODVWWRDOWHUR[\FRGRQHLQGXFHGVXEMHFWLYH
analgesic, and physiological effects. The effects of minocycline on
R[\FRGRQHLQGXFHGVXEMHFWLYHDQDOJHVLFDQGSK\VLRORJLFDOHIIHFWVLQQRQGHSHQGHQWRSLRLGDEXVHUVDOVRZHUHH[DPLQHG7KHUHVXOWV
of these studies suggest that ibudilast reduced opioid withdrawal
V\PSWRPVDQGDFFHQWXDWHGR[\FRGRQHLQGXFHGDQDOJHVLDZLWKRXWDOWHULQJWKHSRVLWLYHVXEMHFWLYHHIIHFWVRIR[\FRGRQH,EXGLODVW
also appeared to produce a dose-dependent reversal of tolerance
WRWKHPLRWLFHIIHFWVRIR[\FRGRQH3UHOLPLQDU\DQDO\VHVRI
the data with minocycline suggest that it produced robust anddose-dependent reductions in the positive subjective effects of
R[\FRGRQHDVZHOODVGUXJFUDYLQJZLWKRXWDOWHULQJLWVUHVSLUDWRU\depressant or miotic effects. The potential utility of these
medications for treating pain and opioid abuse will be discussed.
Disclosures: We gratefully acknowledge the research support
provided by NIDA (DA09236), and the supplemental funding
and provision of ibudilast by MediciNova Inc. In addition, SDC
FRQVXOWHGIRU3À]HUDQG6DOL[GXULQJWKHSDVW\HDUDQGVKHUHFHLYHV
research funding from Reckitt Benckiser and study medicationfrom Omeros. KWJ is an employee of MediciNova. The authors
KDYHQRRWKHUSRWHQWLDOFRQÁLFWVRILQWHUHVWWRGHFODUH

www.dcconferences.com.au/inrc2013/

Departments of Comparative Biology & Experimental Medicine, andPhysiology & Pharmacology, Hotchkiss Brain Institute, University of Calgary,
Calgary, Alberta,T2N 4N1, Canada

Opiates such as morphine are the gold standard pain relieving
drugs, but their use is plagued with debilitating side effects. Twomain problems that limit the treatment of pain with opiates are:SDUDGR[LFDOK\SHUDOJHVLD LQFUHDVHGSDLQVHQVLWLYLW\ DQGWROHUDQFH
(diminished analgesic potency). The prevailing dogma is that bothof these problems are inevitable consequences of opiate use,
DQGWKDWWROHUDQFHDQGK\SHUDOJHVLDUHÁHFWDVLQJOHXQGHUO\LQJ
mechanism. On the contrary, we found that hyperalgesia-inducing
treatment with morphine downregulates the K+-Cl- cotransporter.&&LPSDLULQJ&OH[WUXVLRQDQGWKXVFDXVLQJGLVLQKLELWLRQ
of spinal lamina I pain-transmitting neurons. Restoring Eanion
reversed the morphine-induced hyperalgesia without affecting
tolerance. The hyperalgesia was also reversed by ablating spinal
PLFURJOLDZLWKWKHLPPXQRWR[LQPDFVDSRULQ,QDGGLWLRQ
we found that morphine acting through µ-receptors increased
H[SUHVVLRQRI$73JDWHG3;UHFHSWRUVLQSULPDU\PLFURJOLD
which drives the release of brain-derived neurotrophic factor(BDNF). This release was gated by a µ-receptor independent
mechanism. Blocking BDNF-TrkB signalling preserved Clhomeostasis in lamina I neurons and reversed the hyperalgesia.
Gene-targeted mice in which BDNF was deleted from microglia,
or mice lacking P2X4 receptors, do not develop hyperalgesia to
PRUSKLQHWUHDWPHQW<HWQHLWKHUPRUSKLQHDQDOJHVLDQRUWROHUDQFH
was affected in these animals. In summary, we discovered thatmorphine hyperalgesia, but not tolerance, is mediated by microgliaWRQHXURQVLJQDOOLQJWKDWXOWLPDWHO\LQFUHDVHVQHXURQDOH[FLWDELOLW\E\GLVUXSWLQJLQWUDFHOOXODU&OKRPHRVWDVLV:HLGHQWLÀHGWKDWWKH
core microglia-to-neuron signalling ensemble critically involved
in morphine hyperalgesia is the P2X4 recetor-BDNF-TrkB-KCC2
FDVFDGH&ROOHFWLYHO\RXUÀQGLQJVSURYLGHDQHZPHFKDQLVWLF
framework for understanding morphine hyperalgesia as being
distinct from those of tolerance. This work was supported by
grants from the Canadian Institutes of Health Research (CIHR) andthe Natural Sciences and Engineering Research Council of Canada(NSERC).

17

PROGRAM ABSTRACTS

6<0326,80

23,2,'6,11211(8521$/&(//6
5(*8/$7,21$1'3527(&7,212)7+(HEART*+HDGULFN-3See Hoe, L.E., 3HDUW-1*ULIÀWK+HDOWK,QVWLWXWH*ULIÀWK8QLYHUVLW\6RXWKSRUW4/'$XVWUDOLD

Background: Opioids and opioid receptors (ORs) are importantin cardiac control, and in governing myocardial tolerance toinjurious stressors. The heart produces pro-dynorphin andpro-enkephalin, which are enzymatically processed to active
peptides. Such opioids modulate cardiac inotropy and vasculartone via OR-dependent and -independent mechanisms. ORs also
modify heart-rate and vagal tone, and negatively control cardiac
ß-adrenergic responses. Recent work reveals a role for opioids and
ORs in cardioprotection, limiting cell damage and dysfunction with
LVFKDHPLDRUK\SR[LDH[RJHQRXV25DJRQLVWVLQGXFHSURWHFWLRQ
pre- or post-insult, with endogenous OR ligands contributing toconditioning responses and hibernation.
Findings: We show acute morphine and delta- and kappa-OR
agonists (BW373U86, U50,488) enhance myocardial tolerance toischaemia-reperfusion (I-R) via PI3K/Akt signaling and Katp channel
modulation in rodents. These effects appear to involve MMPdependent EGFR transactivation. Intriguingly, 3- to 5-days of lowlevel morphine or a delta-OR agonist induces a unique protectedstate independent of major protective mediators (eg. NOS, PI3K/
Akt), and that persists for up to a week post-stimulus. Since I-R
occurs primarily in the elderly, and in subjects with common
co-morbidities such as diabetes and hypertension, we tested theeffects of 5-day morphine conditioning in aged (18-24 mth) miceand a murine model of type II diabetes (T2D; 75 mg/kg STZ and 10
weeks of high fat feeding). Aging and T2D both impaired contractile
UHFRYHULHVDQGH[DJJHUDWHGLQIDUFWLRQLQPRXVHKHDUWVVXEMHFWHG
WR,5H[YLYRGD\WUHDWPHQWZLWKVXEFXWDQHRXVPRUSKLQH
(25-75 mg pellets) markedly protected against I-R injury and cell
death in both aged and T2D hearts, whereas acute OR responses
were ablated (as was protection via ischaemic preconditioning).
Protection was associated with improvements in Ca2+-dependent
mitochondrial swelling and respiratory function. Chronic morphinealso reversed T2D features in vivo, improving ventricular contractile
function, countering hyperglycaemia and reducing body weight.
Summary:$FXWHRUFKURQLFDFWLYDWLRQRI25VFDQH[HUW
powerful protective actions in the heart via distinct signallingmechanisms. While effects of acute OR agonism are negated withage and disease, novel protection via chronic morphine persists.
7KLVODWWHUUHVSRQVHDOVRH[KLELWVXQLTXHDQWLGLDEHWLFIHDWXUHVThe OR system warrants further investigation as a means ofmanipulating cardiovascular responses to diabetes and ischaemic
heart disease.

18

International Narcotics Research Conference 2013

PROGRAM ABSTRACTS+27723,&6

0253+,1(02'8/$7(6%5($67&$1&(5&(//0(7$67$7,&327(17,$/
Imani, B.A.1, Baran, J.2&DERW3-1 3DUDW01,2
12School of Pharmacy, University of Queensland,Woolloongabba, Australia
Department of Anesthesia Research, Cleveland Clinic, Cleveland, OH

Appropriate pain management during and after cancer surgery
may play a role in prevention of tumor recurrence and metastasis.
Opioids are proven to be highly effective perioperative analgesics
and are widely used in cancer surgery patients. Using a mousesyngeneic model of breast cancer, we studied the effect ofmorphine on tumor growth and dissemination to lungs. I.P.
injection of morphine (10mg/kg) to mice (n=8) every 12h for
3 consecutive days, caused a reduction in breast tumor growth
and tumor cell dissemination to the lungs as measured 18 days
after tumor inoculation. Morphine treatment also caused a
UHGXFWLRQLQFLUFXODWLQJSURWHRO\WLFHQ]\PHVRIH[WUDFHOOXODUPDWUL[(&0 PDWUL[PHWDOORSURWHLQDVH 003 DQGXURNLQDVHOLNH
plasminogen activator (uPA) measured by gelatin and caseinplasminogen zymography, respectively. Furthermore, we tested
the effect of morphine on co-cultures of breast cancer cellswith stromal cells, either endothelial cells or macrophages. In
co-cultures of 4T1 breast cancer cells with murine RAW264.7
PDFURSKDJHVRUPXULQH+9HQGRWKHOLDOFHOOVWKHOHYHORIPDWUL[proteases produced by cells was increased, and so was the level
RIPDWUL[SURWHDVHLQKLELWRUV7,03V,QWHUHVWLQJO\PRUSKLQH
treatment of these co-cultures reduced the level of MMP-9 andincreased its endogenous inhibitor, TIMP-1, thereby altering theRYHUDOOSURWHRO\WLFSURÀOH0RUSKLQHDIIHFWHGFRFXOWXUHVEXW
not cells grown individually. This suggests that anti-tumor effects
of morphine are mediated through modulation of paracrine
FRPPXQLFDWLRQEHWZHHQFDQFHUFHOOVDQGLQÀOWUDWLQJFHOOV())(&72)'(/7$23,2,'5(&(3725621
+80$1.(5$7,12&<7($'+(6,21$1'
0,*5$7,21'85,1*:281'+($/,1*
%LJOLDUGL4L017HR</1, Neumann, C.1%LJOLDUGL3/1,2
1
Institute of Medical Biology, Agency for Science,Technology & Research(A*STAR), Singapore
2
Department of Internal Medicine, School of Medicine, National University
Hospital, 5 Lower Kent Ridge Road, Singapore 119074

Abbreviations: Beta-1 Integrin , ITGB1; Beta-Catenin, ȕ-Cat;
Delta-opioid receptor, DOR; Desmoglein, DSG; Desmoplakin, DSP;([WUDFHOOXODUVLJQDOUHJXODWHGNLQDVHV(UN,QWHUPHGLDWHÀODPHQWV,)0HW(QNHSKDOLQ0HW(QN1DOWULQGROH1DO2YHUH[SUHVVLRQ2(Wild type, WT; Protein Kinase C, PKC
In addition to its analgesic functions, the peripheral opioid receptor
V\VWHPDIIHFWVVNLQKRPHRVWDVLVE\LQÁXHQFLQJFHOOGLIIHUHQWLDWLRQ
migration, and adhesion. Previous studies have shown that wound
KHDOLQJDQGVNLQKRPHRVWDVLVDUHVLJQLÀFDQWO\DOWHUHGLQGHOWDRSLRLGUHFHSWRUNQRFNRXWPLFH '25 ([WHQGLQJWKHVHREVHUYDWLRQV
we now show that DOR activation in human keratinocytes
regulates intercellular adhesion molecules facilitating cell migration
LQYLWUR'HVPRJOHLQ '6* DQG'6*H[SUHVVLRQZDVXS
regulated in skin from DOR-/- mice, and down-regulated in DORRYHUH[SUHVVLQJ '252( 17(57NHUDWLQRF\WHV7KHH[SUHVVLRQ
pattern of the desmosomal plaque proteins beta-catenin (ȕ-Cat)and desmoplakin (DSP) was also altered, resulting in less stableintercellular adhesion. Consequently, DOR-stimulated migrationwww.dcconferences.com.au/inrc2013/

and wound recovery were enhanced in DOR-OE cells in an in vitro
scratch assay. DOR effects on cell migration and adhesion weremediated by the PKC signalling pathway, and could be antagonized
E\VSHFLÀF3.&Į/ȕ inhibition. Finally, DOR-OE cells build long and
FKDUDFWHULVWLFSURWUXVLRQVFRQWDLQLQJÀODPHQWRXVDFWLQDQG'25
at their free edges, indicating a migratory phenotype. In conclusion,these results from transgenic mice and human immortalized
keratinocyte cell lines show that the opioid receptors affectcell-cell adhesion and wound healing mechanisms, underlining theimportance of skin-nerve interactions in wound healing and skinhomeostasis.

52/(2)23,2,'6<67(0,17+(&(//',))(5(17,$7,21)5203/85,327(1767(0
CELLS*Kuzumaki, N.1,2<DPDPL]X.3,4, Narita, M.1, Okano, H.2,
Narita, M.1
Dept. Pharmacol., Hoshi Univ. Sch. Pharm. Pharmaceut. Sci.,Tokyo, Japan
Dept. Physiol., Keio Univ. Sch. Med.Tokyo, Japan
3
NIH, NIA, Developmental Genomics and Aging Section, Laboratory of
Genetics, MD, USA
4
Laboratory of Stem cell Differentiation, Stem Cell Research Center, Institutefor Frontier Medical Sciences, Kyoto University, Kyoto, Japan1
2

Since the advent of human embryonic stem cells (ESCs), stem
cell research has been developing at a breathtaking pace. The
pluripotent stem cells have the ability to differentiate into
any cell types including into those with therapeutic potential,
after practically unlimited self-renewal in the stem cell state.
The pluripotent stem cells hold enormous promise as tools
for understanding normal development and disease, and just as
importantly, for cell therapy applications to treat devastating andcurrently incurable disorders. However a potential mechanism thatis poorly understood entails the role of endogenous hormones,
JURZWKIDFWRUVDQGQHXURWUDQVPLWWHUVLQLQÁXHQFLQJVWHPFHOO
fate decisions. Therefore, it is important to identify the intrinsic
DQGRUH[WULQVLFIDFWRUVWKDWUHJXODWHWKHXQGHUO\LQJPROHFXODU
mechanisms involved in ESC self-renewal, proliferation anddifferentiation.
+HUHZHSURÀOHGWKHH[SUHVVLRQRIRSLRLGUHODWHGJHQHVLQ
undifferentiated ES cells and differentiated neural stem cells.
We found that delta opioid receptor (DOR), but not mu opioid
receptor (MOR) or kappa opioid receptor (KOR), plays a rolein neurogenesis and neuroprotection in neural stem cells. We
also demonstrated a novel mechanism for the regulation of EC
differentiation and vascular formation through the opioid system.
(VSHFLDOO\.25ZDVKLJKO\H[SUHVVHGLQHPEU\RQLFVWHPFHOO²
derived Flk1+ vascular progenitors. Addition of KOR agonists to
ES cells-derived Flk1+ vascular progenitors inhibited endothelial
cell differentiation, indicating that the KOR system is, thus, a new
UHJXODWRURIYDVFXODUGHYHORSPHQWWKDWVLPXOWDQHRXVO\PRGLÀHV(&
GLIIHUHQWLDWLRQDQGYDVFXODUSDWKÀQGLQJ:HFRQFOXGHWKDWRSLRLGVmay be responsible for ES cells differentiation at the developmental
stage.

19

PROGRAM ABSTRACTS

6<0326,80

0($685(0(17$1'0(&+$1,602)/,*$1'%,$6$77+(23,2,'5(&(3725
Kelly, E.
School of Physiology and Pharmacology, University of Bristol, Bristol BS8 1TD,UK

Drugs acting at the µ opioid receptor (MOPr) are amongst themost important therapeutic agents available, but much remains
to be understood about the molecular signals that arise following
activation of this G protein coupled receptor by opioid agonists. It
is likely that signalling via both G protein- and arrestin-dependent
pathways is important for MOPr function. However, the relative
abilities of MOPr agonists to signal via these two pathways may
not be the same for all agonists, a phenomenon termed ligand bias
or functional selectivity. By identifying MOPr ligands that display
G protein or arrestin bias at MOPr, it may be possible to develop
GUXJVZKLFKKDYHDEHWWHUWKHUDSHXWLFSURÀOHZLWKIRUH[DPSOH
fewer adverse effects, less liability to promote tolerance toanalgesic effects, or less likelihood to induce dependence.
In this presentation I will discuss how cellular assays for G
protein- and arrestin-dependent signalling pathways can be used todetermine the presence of ligand bias for a series of MOPr ligands.
I will discuss how the Operational Model of Agonism, developed
by Black and Leff over a quarter of a century ago, is becoming an
increasingly important means to analyse agonism and detect ligand
bias. In addition, I will discuss potential mechanisms that underlie
ligand bias at MOPr, such as ligand-dependent phosphorylationof multiple residues in the COOH-terminus of the receptor.
Finally, I will discuss the possible advantages and disadvantages ofdeveloping ligands with either G protein or arrestin bias at MOPr,and indeed whether or not we know enough about the MOPrsignalling pathways in brain to actually know what sort of bias may
be preferable.

/,*$1'$1'&(//'(3(1'(17'(7(50,1$1762)'(/7$23,2,'5(&(3725
'25 /,*$1'6,*1$//,1*
3LQH\UR*
Department of Psychiatry and Department of Pharmacology, University ofMontreal. Ste-Justine Hospital Research Center. Montreal, Quebec, Canada

Opioid receptor agonists have potent analgesic actions but theseeffects are restricted by the development of tolerance. However,tolerance is not the same for all ligands and these differences
have been attributed, at least in part, to biased agonism. This
notion is based on the observation that ligands with similar G
protein-mediated responses may display distinct potential for
WROHUDQFHGHSHQGLQJRQWKHLULQWHUQDOL]DWLRQSURÀOHV$SUREOHP
with this notion is that some of these observations have relied
RQFRPSDULVRQRI(PD[YDOXHVUDWKHUWKDQHVWLPDWLRQRIUHODWLYHHIÀFDFLHV+HUHZHVRXJKWWRPHDVXUHZKHWKHU'25DJRQLVWV
displayed bias in cyclase and internalization responses. To do so,
we used the operational model of Black and Leff to generate
WUDQVGXFWLRQFRHIÀFLHQWV ORJWDX.$ IRUHDFKRIWKHUHVSRQVHVRILQWHUHVW7KHVHFRHIÀFLHQWVFDSWXUHGLIIHUHQFHVERWKLQOLJDQGHIÀFDF\ WDX DQG´IXQFWLRQDODIÀQLW\µ .$ ZKLFKLQWXLWLYHO\FRUUHVSRQGVWROLJDQGDIÀQLW\IRUWKHUHFHSWRUVWDWH V PHGLDWLQJWKHUHVSRQVHRILQWHUHVW&RPSDULVRQRIWUDQVGXFWLRQFRHIÀFLHQWV
obtained in HEK293 cells indicated that TIPP and UFP512 wereUHVSHFWLYHO\DQGIROGPRUHHIÀFLHQWLQF\FODVHWKDQinternalization assays as compared to DPDPE. SNC-80 was
VLPLODUO\ELDVHGEHLQJIROGPRUHHIÀFLHQWLQF\FODVHWKDQinternalization assays.
Importantly, molecular determinants of internalization were
not the same in HEK293 cells and neurons, raising the questionZKHWKHUELDVLGHQWLÀHGLQRQHFHOOW\SHFRXOGEHUHOHYDQWWRWKH
other. In particular, ȕarrestins contributed to DOR internalizationin the two cellular backgrounds, but PKC and GRK2 (G proteincoupled receptor kinase 2) were only involved in neurons. These
GLIIHUHQFHVUHVXOWHGLQKLJKHUUHODWLYH(PD[LQWHUQDOL]DWLRQYDOXHV
for TIPP and UFP512 but not for SNC-80 (DPDPE was the
VWDQGDUG 0RUHRYHUWKHH[WHQWWRZKLFKGLIIHUHQWOLJDQGVUHOLHG
on ȕDUUVDQG3.&VWRLQGXFHPD[LPDOLQWHUQDOL]DWLRQLQQHXURQV
ZHUHERWKFRUUHODWHGZLWKWUDQVGXFWLRQFRHIÀFLHQWVIRUF\FODVH
inhibition in HEK293 cells, implying a common substrate for these
responses that could be evidenced across cellular backgrounds.
GRK2 contribution to ligand-induced internalization was unrelated
to the other signals, suggesting that the activity of this kinase could
determine internalization bias in neurons.
Taken together these observations indicate that for bias to be
independent of cellular background, it should be measured on
actual signals and not phenotypic cellular responses.

20International Narcotics Research Conference 2013

PROGRAM ABSTRACTS+27723,&6

48$17,),&$7,212))81&7,21$/
6(/(&7,9,7<2)(1'2*(128623,2,'6$7
7+(0823,2,'5(&(3725

IDENTIFICATION OF ALLOSTERIC
02'8/$72562)23,2,'5(&(37256%<+,*+7+528*+3876&5((1,1*Thompson, G., Christopoulos, A. and *Canals, M.

Burford, N.T.1, Wehrman,T.S.2, Clark, M.J.3, Gerritz, S.W.1,
Banks, M.1, O’Connell, J.1,Traynor, J.R.3 and *Alt, A.1

Monash Institute of Pharmaceutical Sciences, Drug Discovery Biology
Laboratory, Parkville,Victoria, Australia.

Functional selectivity, or biased agonism, refers to the paradigm
whereby different ligands, binding to the same receptor, engender
distinct receptor conformations linked to divergent signalling
pathways and physiological outcomes. The description of numerous
functionally selective ligands in the last decade has prompted theidea that functional selectivity represents a novel direction for drug
discovery whereby drugs can be “pathway selective” in addition to´UHFHSWRUVHOHFWLYHµ$VVXFKELDVHGOLJDQGVPD\DOORZIRUWKHÀQHtuning of receptor signalling towards desired pathways and awayfrom unwanted signalling cascades such as those mediating adverse
side effects. However, functional selectivity has broader implications
for GPCR-directed therapeutics when considering systems
in which multiple endogenous ligands target a given receptor,
as it raises the possibility of “endogenous (natural) functional
VHOHFWLYLW\µPHGLDWLQJVSHFLÀFSK\VLRORJLFDOUHVSRQVHV8QIRUWXQDWHO\ZLWKIHZH[FHSWLRQVPRVWGHVFULSWLRQVRIELDVHG
agonism have been based on qualitative comparisons between
ligands, and the number of studies quantifying bias in a statisticallyrobust manner is still very low. However, in order to apply
functional selectivity therapeutically and in the most effective
manner, it is necessary to incorporate parameters that describe
the degree of bias in a manner that can inform structure activity
studies and drug candidate selection matrices. We have developed
an analytical method, based on the operational model of agonism,that can be applied to standard concentration-response data toderive “bias factors” for each agonist. These bias factors can beevaluated statistically in a system-independent manner.
7KHFXUUHQWVWXG\DSSOLHGWKLVPHWKRGWRWKHTXDQWLÀFDWLRQRI
functional selectivity of endogenous opioids at the mu-opioid
receptor (MOR) in cellular model systems. This has allowed
WKHJHQHUDWLRQRIWKHÀUVWDFWLYLW\SURÀOHVIRUWKHHQGRJHQRXVRSLRLGIDPLO\DQGWKHGHWHFWLRQRIGLYHUJHQWVLJQDOÀQJHUSULQWV
for this class of ligands. Unravelling unappreciated activity
patterns of endogenous opioid peptides can provide essential
information regarding the mechanisms behind opioid-induced
physiological responses (from analgesia to respiratory depression
DQGFRQVWLSDWLRQ WKDWFDQWKHQEHH[SORLWHGWRGHVLJQQRYHOGUXJ
targets with effective analgesia but with minimal side effects. As
VXFKIXWXUHVWXGLHVZLOODGGUHVVWKHSRWHQWLDOIRUWKHELDVSURÀOHVto be predictive in physiological systems and to further validate
functional selectivity in vivo.

www.dcconferences.com.au/inrc2013/

1
Molecular Sciences and Candidate Optimization, Bristol-Myers Squibb,
Wallingford, CT, USA, 2 DiscoveRx Corporation, Fremont, CA, USA and 3
Department of Pharmacology, University of Michigan, Ann Arbor, MI, USA.

Allosteric modulators of G protein-coupled receptors have anumber of potential advantages over traditional orthostericagonists and antagonists, including potential improvement in
WROHUDQFHVDIHW\DQGVLGHHIIHFWSURÀOHVA high-throughput screen (HTS) of >1 million compounds
was designed to identify positive allosteric modulators (PAMs)of opioid receptors. The HTS measured the enhancement ofendomorphin-I and Leu-enkephalin-mediated recruitment ofȕDUUHVWLQLQ86KXPDQRVWHRVDUFRPDFHOOVH[SUHVVLQJP- and/or Gopioid receptors; the assay was designed to simultaneously
identify PAMs of P- and/or G opioid receptors in a single screen.
The HTS used an N-terminal deletion mutant of ȕ-galactosidase
IXVHGWRWKH&WHUPLQXVRIVWDEO\H[SUHVVHGȕ-arrestin-2 anda mutated amino-terminal fragment of ȕ-galactosidase fused
WRWKHFDUER[\OWHUPLQXVRIWKHDSSURSULDWHRSLRLGUHFHSWRU
Binding of ȕ-arrestin-2 to activated opioid receptor results in acomplementation of the enzyme and reconstitution of enzyme
DFWLYLW\)URPWKH+76VFUHHQZHLGHQWLÀHGWZRVHOHFWLYHRSLRLG
receptor PAMs: (E)-N(-2,6-dichlorophenyl)-4-(4-nitrophenyl)2,3-dihydro-1,3-thiazol-2-imine (BMS-986121) and 2-(3-bromoPHWKR[\SKHQ\O  FKORUREHQ]HQHVXOIRQ\O WKLD]ROLGLQH
(BMS-986122). In vitro, both compounds produced concentrationdependent and saturable leftward shifts in concentration-response
curves for the µ-opioid receptor agonist endomorphin-I torecruit ȕDUUHVWLQSURGXFLQJDQDSSUR[LPDWHIROGPD[LPDO
leftward shift in agonist response, with Kb (shift50) values of 2-5µM. While BMS-986121 behaved as a “pure” PAM, BMS-986122
displayed allosteric agonist activity, behaving as a PAM at lower
FRQFHQWUDWLRQVEXWH[KLELWLQJLWVRZQLQWULQVLFDJRQLVWDFWLYLW\DWhigher concentrations.
Structure-activity relationships within the thiazolidine chemotype
ZHUHH[SORUHGDQGWKHHIIHFWVRIFKHPLFDOPRGLÀFDWLRQVWR
BMS-986122 on µ-PAM activity will be described; however none
showed improved PAM activity over the original hit. While BMS986122 was seen to be selective for the µ-opioid receptor, some
DQDORJVZHUHIRXQGWRH[KLELWZHDN3$0DFWLYLW\DWG-opioid
UHFHSWRUV$GGLWLRQDOO\WZRDQDORJVZHUHLGHQWLÀHGDV´VLOHQWµallosteric modulators (SAMs) of the µ-opioid receptor and acted
as competitive antagonists for the allosteric site to which BMSELQGVFRQÀUPLQJWKHSUHVHQFHRIDGLVFUHWHDOORVWHULF
binding site on the µ opioid receptor.

21

PROGRAM ABSTRACTS+27723,&6

326,7,9($//267(5,&02'8/$7,212)7+(0823,2,'5(&(3725
Clark, M.J.1, Burford, N.T.2, Livingston, K.E.1, Mahoney, J.1,
Alt, A.2, *Traynor, J.R.1
Department of Pharmacology, University of Michigan, Ann Arbor, MI, USA,
Molecular Sciences and Candidate Optimization, Bristol-Myers Squibb,
Wallingford, CT, USA
1
2

Allosteric ligands for G protein coupled receptors (GPCRs)bind to a site on the receptor that is distinct from the site thatbinds orthosteric (or endogenous) agonists. Positive allosteric
PRGXODWRUV 3$0V JHQHUDOO\ODFNLQWULQVLFHIÀFDF\EXWLQFUHDVH
WKHELQGLQJDIÀQLW\DQGRUHIÀFDF\RIRUWKRVWHULFDJRQLVWV7KLV
enhancement may be agonist- and signaling pathway-dependent.
Thus, PAMs active at the mu-opioid receptor (MOR-PAMs) could
promote activation of select signaling pathways, thereby potentially
VHSDUDWLQJEHQHÀFLDODQGXQZDQWHGHIIHFWV,QDGGLWLRQ0253$0V
might augment the action of endogenous opioid peptides and so
avoid compensatory mechanisms that are triggered by sustained
UHFHSWRUDFWLYDWLRQSURGXFHGE\H[RJHQRXVRSLRLGV+HUHZH
report an initial characterization of a MOR-PAM, BMS-986122,
LGHQWLÀHGE\DKLJKWKURXJKSXWVFUHHQ%06LPSURYHG
WKHELQGLQJDIÀQLW\RIWKH025DJRQLVW'$0*2E\IROGLQ&FHOOVH[SUHVVLQJ025 &025 ZLWKRXWDOWHULQJELQGLQJRIWKH
antagonist diprenorphine. The potency of DAMGO to stimulate[35S]GTPJS binding in C6MOR cells and mouse brain membranes
was increased by 4-7-fold in the presence of BMS-986122. With
the partial agonists morphine and endomorphin-1 there was a
3-fold increase in potency together with a 1.7-fold increase in
PD[LPDO>6@*73JS stimulation. Thus, BMS-986122 is both anDIÀQLW\DQGDQHIÀFDF\PRGXODWRU7KHDIÀQLW\RI%06
FDOFXODWHGXVLQJDQDOORVWHULFWHUQDU\FRPSOH[PRGHOZDVPM.
A close analog of BMS-986122 acted as a competitive antagonistof the PAM, but not diprenorphine, indicating the presence of a
GLVWLQFWDOORVWHULFELQGLQJVLWH7RFRQÀUPWKHDOORVWHULFELQGLQJ
VLWHLVRQ025ZHXVHGSXULÀHG025SURWHLQLQFRPSOH[ZLWK*SURWHLQKHWHURWULPHUDQG*'37KLVFRPSOH[ZDVLQFRUSRUDWHG
into a phospholipid bilayer in the form of recombinant high density
OLSRSURWHLQSDUWLFOHV,QWKLVSUHSDUDWLRQRQO\DVLQJOHORZDIÀQLW\low DAMGO binding site (Ki = 500nM) was observed. BMSFDXVHGWKHDSSHDUDQFHRIDKLJKDIÀQLW\VLWHIRU'$0*2
(Ki = 20nM) that was lost in the presence of unlabelled GTPJS.
These studies provide proof-of-concept for the development ofnovel allosteric modulators of MOR that may have therapeuticpotential in improving pain management. Supported in part by
MH083754.

22(1'20253+,1$1$/2*6:,7+5('8&('$%86(/,$%,/,7<5(63,5$725<'(35(66,21
02725,03$,50(1772/(5$1&($1'*/,$/
$&7,9$7,215(/$7,9(720253+,1(=DGLQD-(1,2,3,4, Nilges, M.R.4, Morgenweck, J.2=KDQJ;1,2,
Hackler, L.1,2, Fasold, M.B.1,2
1
SE Louisiana Veterans HCS, NEW ORLEANS, LA and Depts of 2 Medicine, 3
Pharmacol., 4 Neurosci. Program,Tulane Univ. Sch. of Med., New Orleans, LA,USA

Opioids acting at the mu opioid receptor (MOR) are the mosteffective analgesics, but adverse side effects severely limit theiruse. Abuse liability results in major medical, societal, and economic
problems, respiratory depression is the cause of death fromoverdose, and tolerance complicates treatment and increases therisk of side effects. Motor and cognitive impairment are especiallyproblematic for older adults. Morphine-induced glial activation iswell characterized in tolerance and may contribute to additional
side effects. The endomorphins (EMs) are potent and selective
agonists at the mu opioid receptor. Three metabolically stable
EM analogs were synthesized and tested for their antinociceptive
DQGVLGHHIIHFWSURÀOHVLQWKHUDW,QUHFHSWRUDQG*73JS assays,
WKHDQDORJVVKRZHGKLJKDIÀQLW\VHOHFWLYLW\DQGHIÀFDF\DW025
relative to delta and kappa opioid receptors. In tests of reward,
PRUSKLQHLQGXFHGVLJQLÀFDQWFRQGLWLRQHGSODFHSUHIHUHQFHDQG
self-administration, while the analogs did not, consistent with
reduced abuse potential. At equiantinociceptive doses, respiratory
GHSUHVVLRQZDVVLJQLÀFDQWO\UHGXFHGIRUDOODQDORJVUHODWLYHWRPRUSKLQH$QDORJGLGQRWLQGXFHVLJQLÀFDQWUHVSLUDWRU\GHSUHVVLRQDWDGRVHSURGXFLQJVLJQLÀFDQWO\ORQJHUDQWLQRFLFHSWLRQ
WKDQPRUSKLQH0RWRULPSDLUPHQWRQDURWRURGZDVVLJQLÀFDQWO\OHVVDQGDQWLQRFLFHSWLRQVLJQLÀFDQWO\ORQJHUDIWHUDQDORJWKDQ
after morphine. Tolerance was determined with cumulative dosing
before and after intrathecal administration of morphine or analogs
for 7 days with an osmotic minipump. Analogs were initially 30-foldmore potent than morphine and, compared to a 61-fold shift in thedose-response curve by morphine, the average shift for the analogs
ZDVRQO\IROGFRQVLVWHQWZLWKVLJQLÀFDQWO\OHVVWROHUDQFH
0RUSKLQHSURGXFHGVLJQLÀFDQWJOLDODFWLYDWLRQDVPHDVXUHGE\increased GFAP, Iba1 and pp38 staining, while the analogs did not.
,QVXPPDU\RXUQRYHO(0DQDORJVVKRZDKLJKO\IDYRUDEOHSURÀOH
relative to morphine as potent analgesics with reduced 1) reward/
abuse liability, 2) respiratory depression, 3) motor impairment,4) tolerance and 5) glial activation. Funded by the VA, ONR, and
DOD.

International Narcotics Research Conference 2013

PROGRAM ABSTRACTS+27723,&6

/,*$1'%,$6('0(&+$1,6062)23,2,'
$17,12&,&(37,21*Bobeck, E. N.1, Ingram, S. L.2, & Morgan, M. M.1
Dept. of Psychology,Washington State University,Vancouver,WA, USA
Dept. of Neurological Surgery, Oregon Health & Science University,
Portland, OR, USA
1
2

Opioids such as morphine bind and activate the mu-opioid
receptor (MOPr) which leads to activation of G-proteins.
Inhibitory G-proteins have been shown to be important in
DQWLQRFLFHSWLRQ,QKLELWLRQRIWKHVH*SURWHLQVZLWKSHUWXVVLVWR[LQ(PTX) causes a greater reduction in morphine antinociception
than that produced by other MOPr agonists. MOPr agonists
also differ in their ability to recruit E-arrestin and induce MOPrinternalization. Morphine produces little internalization, whereas
DAMGO and fentanyl produce rapid internalization. Althoughmuch is known about these ligand-biased signaling pathways,
OLWWOHLVNQRZQDERXWWKHEHKDYLRUDOVLJQLÀFDQFHRIDFWLYDWLRQRIWKHVHVLJQDOLQJSDWKZD\V7KHSXUSRVHRIWKLVH[SHULPHQWZDV
to test the hypothesis that the signaling pathway (i.e., G-protein
and internalization) that contributes to antinociception differs
depending on whether the MOPr is bound by morphine, DAMGO,
or fentanyl. All drugs will be injected into the ventrolateral
SHULDTXHGXFWDOJUD\ YO3$* VRDVWROLQNVSHFLÀFVLJQDOLQJ
pathways to antinociception. To evaluate the role of G-proteins in
antinociception, PTX (5 or 50 ng/0.5 µL) was microinjected into
the vlPAG prior to cumulative doses of the opioid. Pretreatment
with 5 or 50 ng PTX caused a rightward shift in the morphinedose-response curve. However, only 50 ng PTX attenuatedantinociception to DAMGO, and neither dose altered fentanylinduced antinociception. To investigate the role of internalizationin antinociception dynamin-dominant negative peptide (dyn-DN;
2 µg/0.5 µL) was administered into vlPAG to prevent receptor
internalization. Pretreatment with dyn-DN did not alter themorphine dose-response curve, but did cause a rightward shift
in the DAMGO dose-response curve and a leftward shift in thefentanyl dose-response curve. These data indicate that morphineuses a G-protein mediated process to induce antinociception,
whereas DAMGO uses both G-protein and dynamin-dependent
mechanisms. The mechanism underlying fentanyl antinociception
is unclear. However, these studies suggest that receptor
internalization may terminate the MOPr signaling that causes
antinociception. These results suggest that antinociception can becaused by distinct mechanisms depending on the agonist. Funding
by NIH grant DA 015498.

www.dcconferences.com.au/inrc2013/23

PROGRAM ABSTRACTS

6<0326,80

(1'2*(128623,2,'6,17+($0<*'$/$
Bagley, E.
)HDUDQGDQ[LHW\DUHDGDSWLYHUHVSRQVHVWKDWDOORZXVWRGHIHQG
ourselves against harm. Neural circuits in the amygdala are key
mediators of fear memory acquisition and storage, but also
UHGXFHWKHIHDUUHVSRQVHZKHQWKHWKUHDWKDVSDVVHG H[WLQFWLRQ 
Disturbance of these circuits is thought to underlie a range of
DQ[LHW\GLVRUGHUVLQKXPDQVLQFOXGLQJSKRELDV376'DQGSDQLFGLVRUGHUV2QHSRVVLEOHPRGXODWRURIH[WLQFWLRQLVWKHHQGRJHQRXV
opioid system, as deletion of enkephalin produces mice that are
KLJKO\DQ[LRXVDQGIHDUIXO7KHH[WLQFWLRQRIWKHIHDUUHVSRQVH
relies on greater activity of a group of GABAergic interneurons
in the amygdala, the intercalated cells inhibiting the fear response.
,QWHUFDODWHGQHXURQVH[SUHVVYHU\KLJKOHYHOVRIHQNHSKDOLQDQGWKH
µ-opiate receptor, strongly suggesting that intercalated neurons
UHOHDVHHQNHSKDOLQWKDWDFWVORFDOO\WRUHJXODWHFHOOXODUH[FLWDELOLW\DQGXOWLPDWHO\DOWHUVH[SUHVVLRQRIWKHIHDUUHVSRQVH2XUGDWD
shows that in the amygdala endogenously released enkephalin
DFWVLQVHYHUDOGLIIHUHQWZD\VWRPRGLI\QHXURQDOH[FLWDELOLW\DQG
synaptic transmission. These actions of the endogenous opioids
occur with minimal stimulation indicating that these peptides
DUHVLJQLÀFDQWPRGXODWRUVRIDP\JGDODIXQFWLRQDQGDUHOLNHO\WR
modulate the fear response. This work may ultimately provide
insights into the physiological role of enkephalin in the intercalated
FHOOVRIWKHDP\JGDODHVSHFLDOO\GXULQJIHDUH[WLQFWLRQDQGSURYLGH
WKHEDVLVIRUEHWWHUWKHUDSHXWLFDSSURDFKHVWRDQ[LHW\GLVRUGHUV

23,2,'3(37,'(6,1'8&(/21*7(50'(35(66,21$7*/87$0$7(5*,&6<1$36(6,17+('256$/675,$780
Atwood, B.K., Kupferschmidt, D.A., ;LRQJ:DQGLovinger,
D.M.
National Institute on Alcohol Abuse and Alcoholism, Rockville, MD, USA

As prescription opioid analgesic abuse rates rise, so does the needWRXQGHUVWDQGWKHORQJWHUPHIIHFWVRIRSLRLGH[SRVXUHRQEUDLQ
function. The dorsal striatum, with roles in skill learning and habit
formation, is a site of importance for opioid-induced neuronalplasticity. Opioid peptides, their target receptors and the peptidase
enzymes that are responsible for terminating their actions are
DEXQGDQWO\H[SUHVVHGLQWKLVEUDLQUHJLRQ+RZHYHUWKHUHLVD
paucity of data concerning opioid roles in long-term synaptic
plasticity in the dorsal striatum. We utilized whole-cell patch clamp
electrophysiological recordings of medium spiny neurons in thedorsal striatum in an acute brain slice preparation to determine
whether opioids could produce long-lasting changes in synaptic
SODVWLFLW\DWH[FLWDWRU\V\QDSVHV:HGLVFRYHUHGWKDWRSLRLG
agonist-induced inhibition is long-lasting and is able to be blocked,
but not reversed, by opioid receptor antagonists. We refer to this
ORQJODVWLQJVXSSUHVVLRQRIH[FLWDWRU\QHXURWUDQVPLVVLRQDVRSLRLG
peptide-induced long-term depression (OP-LTD). We also foundthat endongeous opioid peptides, released locally in the striatum,
could produce OP-LTD. Mu and delta opioid-mediated forms of
OP-LTD are dissociable, in that they summate, differentially occlude
LTD mediated by endocannabinoids, and are affected differently
E\LQYLYRH[SRVXUHWRWKHRSLRLGDQDOJHVLFR[\FRGRQH7KHVH
data demonstrate a novel form of long-lasting synaptic plasticity in
WKHGRUVDOVWULDWXPWKDWLVLQGXFHGE\DEULHIH[SRVXUHWRRSLRLG
peptides.
This research was supported by the Division of Intramural Clinical and Basic
Research of the National Institute on Alcohol Abuse and Alcoholism, US National
Institutes of Health.

24International Narcotics Research Conference 2013

PROGRAM ABSTRACTS+27723,&6

5(3($7('$9(56,9(675(66(;32685(5('8&(60823,2,'5(&(3725(;35(66,21
,17+(,17(5&$/$7('18&/(,2)7+($0<*'$/$2)5$76*Cox, B.M., Silveira, J.T., Gouty, S. and Cote,T.E.
Dept. of Pharmacology, Uniformed Services University, Bethesda MD 20814,
USA

0XUHFHSWRUV 023U DUHVWURQJO\H[SUHVVHGLQWKHFHOOFOXVWHUV
of the intercalated nuclei (ITC) that surround the lateral (LA)and basolateral (BLA) nuclei in rat amygdala (Likhtik et al, 2008).
ITC neurons may play a role in regulating activity in the neuronalconnections from the BLA, which mediates the integration of
FRQGLWLRQHGDQGXQFRQGLWLRQHGQR[LRXVVWLPXOLWRWKHFHQWUDO
amygdaloid nuclei (CeA), the major output nucleus of the amygdala
regulating motor and autonomic responses (Pape & Pare, 2010).
We hypothesized that activation of amygdaloid pathways by
H[SRVXUHWRUHSHDWHGDYHUVLYHVWLPXOLPLJKWFKDQJHWKHH[WHQWRIDFWLYDWLRQRUWKHOHYHORIH[SUHVVLRQRI023ULQWKH,7&:H
therefore determined the effects of repeated aversive stress in
UDWVRQWKHH[SUHVVLRQRI023ULQWKH,7&DQGRQRSLRLGSHSWLGH
H[SUHVVLRQLQDP\JGDODQXFOHLXVLQJDVWUHVVIXOVWLPXOXV UHVWUDLQW
for 2 hr with intermittent unpredictable tail shock, repeated on
WKUHHFRQVHFXWLYHGD\V WKDWKDVEHHQXVHGDVDQH[SHULPHQWDO
model for post-traumatic stress disorder (Manion et al, 2007).
$WKUDQGKUDIWHUWKHHQGRIWKHWKLUGVWUHVVH[SRVXUHWKHOHYHORI023UH[SUHVVLRQZDVGHWHUPLQHGE\ÁXRUHVFHQFH
immuno-histochemistry in individual ITC cell groups at three A-P
levels through the amygdala. The level of MOPr in ITC nuclei was
UHGXFHGE\DERXWDWKUDQGE\DERXWDWKUDIWHU
WKHWKLUGGD\RIVWUHVVH[SRVXUH(QGRJHQRXVRSLRLGVDUHDOVR
H[SUHVVHGLQDP\JGDODZLWKWKHKLJKHVWOHYHOVRIH[SUHVVLRQLQWKH
&H$1RVLJQLÀFDQWVWUHVVUHODWHGFKDQJHVLQWKHH[SUHVVLRQRISURHQNHSKDOLQP51$ZHUHREVHUYHGEXWWKHUHZDVDVLJQLÀFDQWLQFUHDVHLQWKHH[SUHVVLRQRISURG\QRUSKLQP51$DQGRI
dynorphin A(1-8) immunoreactivity (DynA-ir) in the paracapsular
VXEGLYLVLRQRIWKH&H$'\Q$LUÀEHUVZHUHDOVRREVHUYHGLQFORVH
SUR[LPLW\ZLWKVRPH,7&FHOOFOXVWHUV7KHVHUHVXOWVVXJJHVWWKDWMOPr in ITC are down-regulated after repeated aversive stress.
It is possible that peptide products of the prodynorphin geneparticipate in this regulation. Further study is required to elucidate
the role of MOPr in ITC in response to aversive stimuli. Supported
by grant PT-73729 from the US Dept of Defense.

www.dcconferences.com.au/inrc2013/

23,2,',1+,%,7,212)*$%$,13876)520
507*$1',17(51(85216,17+(97$$)7(5&+521,&0253+,1(75($70(17Matsui, A.1, Jarvie, B.C.2, Hentges, S.T.2, Williams, J.T.1
1
Vollum Institute, Oregon Health and Science University, Portland, OR, 97239,
USA
2
Department of Biomedical Sciences, Colorado State University, Fort Collins,
CO, 80523, USA

The activity of dopamine neurons is modulated by the balance
EHWZHHQH[FLWDWRU\DQGLQKLELWRU\LQSXWV2SLRLGVLQFUHDVH
dopamine neuron activity through inhibition of GABA input
to dopamine neurons. The recent discovery of a dense opioidsensitive GABA inputs originating from the rostromedial
tegmental nucleus (RMTg) brings into question the role of GABA
interneruons in the ventral tegmental area (VTA). The present
VWXG\XWLOL]HGDQRSWRJHQHWLFVLQUDWVWRH[DPLQHDUHODWLYH
opioid sensitivity of GABA inputs from interneurons in VTA andRMTg neurons. The results show a relative insensitivity of VTA
interneuron input to opioids. When animals were treated withmorphine continuously for 1 week, the inhibition of GABA-A
IPSCs by MOR agonists was reduced, indicating development oftolerance particularly the GABA input from RMTg. This study
demonstrates that GABA inputs from VTA interneurons were lesssensitive to opioids compared to RMTg neurons and less toleranceof this pathway was observed. The functional connection fromthe RMTg to the VTA is the key structure in the MOR dependent
regulation of dopamine neurons. Supported by DA08163,
DA04523 (JTW).

25

PROGRAM ABSTRACTS

6<0326,80

52/($1'5(*8/$7,212)7+('(/7$23,2,'
5(&(3725,13$,1
Gendron, L.
Department of Physiology and Biophysics, Université de Sherbrooke,
Sherbrooke, Quebec, Canada:HKDYHSUHYLRXVO\VKRZQWKDWWKHDQDOJHVLFHIÀFDF\RIGHOWD
opioid receptor (DOPr) agonists can be enhanced by prolonged
PRUSKLQHWUHDWPHQWDQGE\FKURQLFLQÁDPPDWRU\SDLQ2YHU
the years, we have demonstrated that the mu opioid receptor
023U LVDQLPSRUWDQWUHJXODWRURIWKHPHPEUDQHH[SUHVVLRQ
and of the functions of DOPr in spinal cord and dorsal root ganglia(DRG) neurons. The mechanisms involved in this regulation are
still unclear but heterodimerization between MOPr and DOPr
was postulated to play an important role in this phenomenon.
Admittedly, to physically interact with each other these receptors
KDYHWREHFRH[SUHVVHGLQWKHVDPHFHOOV7KLVKRZHYHUUHPDLQV
highly controversial. Indeed, in spinal cord and DRG, MOPr and
DOPr were found to be segregated in different population of cells,
at least in the mouse. This segregation between MOPr and DOPr
LVIXUWKHUVXSSRUWHGE\UHFHQWÀQGLQJVVKRZLQJWKDWDFWLYDWLRQRI
023UDQG'23UVSHFLÀFDOO\LQKLELWVGLVWLQFWSDLQPRGDOLWLHV7R
address this controversy and to determine whether or not MOPr
DQG'23UDUHH[SUHVVHGLQVXEVWDQFH3FRQWDLQLQJQHXURQVZH
have combined behavioral tests, in vivo electrophysiology andimmunolabeling and found that both spinal MOPr and DOPr
DWWHQXDWHQR[LRXVEHKDYLRUDQGLQKLELWVXEVWDQFH3UHOHDVHLQGXFHG
by intraplantar formalin and capsaicin. We also found that activation
RIWKHVHUHFHSWRUVSRWHQWO\LQKLELWVGLIIXVHQR[LRXVLQKLELWRU\FRQWUROV '1,& LQGXFHGE\WKHUPDODQGPHFKDQLFDOQR[LRXV
stimuli applied to the hindpaw, an effect also accompanied by an
inhibition of substance P release in the spinal cord. Our results
WKHUHIRUHVXJJHVWWKDW023UDQG'23UDUHERWKH[SUHVVHG
in a subpopulation of primary afferents containing substance P
where physical interactions between them might participate in
WKHUHJXODWLRQRI'23U0RVWLPSRUWDQWO\RXUÀQGLQJVLQGLFDWH
that activation of spinal MOPr and DOPr similarly inhibits DNIC
HYRNHGHLWKHUE\QR[LRXVWKHUPDORUPHFKDQLFDOVWLPXOL7KH
VSHFLÀFUROHRIWKH023U'23UKHWHURGLPHULQWKHUHJXODWLRQRI
'23UWUDIÀFNLQJUHPDLQVWREHDGGUHVVHG
This work was supported by the FRQ-S, the CIHR and the NSERC. LG is therecipient of a J1 salary support from the Fonds de la Recherche du Québec –Santé.

*3527(,1&283/('5(&(3725668&+$6
7+('(/7$23,2,'$1'25/5(&(37256
$5($%/(7202'8/$7($*21,676,*1$/,1*
VIA A ROCK, LIMK AND E$55(67,13$7+:$<Mittal, N.1, Roberts, K.13DO.2, Bentolila, L.A.3,4, Cahill, C.5,
3UDGKDQ$1, Conner, D.6, DeFea, K.2, Evans, C.1, Walwyn,
W.1
1
Dept. of Psychiatry and Biobehavioral Sciences, Stefan Hatos Center
for Neuropharmacology, Semel Institute, UCLA, 3Dept of Chemistry andBiochemistry, 4California NanoSystems Institute, UCLA, 2Div. of Biomedical
6FLHQFHV&HOO0ROHFXODUDQG'HYHORSPHQWDO%LRORJ\3URJUDP8&55Deptof Anesthesia and Perioperative Care, UCI, CA. 6Dept of Genetics, Harvard
Medical School, Boston, MA

such signaling. This mechanism allows agonists of the delta opioid
UHFHSWRURU2SLRLG5HFHSWRU/LNH 25/ WRDFWLYDWHFRÀOLQDQ
actin modulating protein, through ROCK and LIMK in a E-arrestin
1 dependent manner. This controls actin polymerization andUHJXODWHVUHFHSWRUH[SRUWIURPWKH*ROJLWRWKHFHOOPHPEUDQH
Receptor function, as determined by receptor inhibition of voltagedependent Ca2+ channels in DRG neurons, and the behavioral
effects of delta opioid receptor and ORL1 agonists are bothLQÁXHQFHGE\WKHVHPHFKDQLVPV)XUWKHUPRUHWKHXSUHJXODWLRQRIGHOWDRSLRLGUHFHSWRUIXQFWLRQIROORZLQJDFKURQLFLQÁDPPDWRU\
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insight into THE mechanisms by which GPCRs signal.

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0$785$7,21
3HWlMl5HSR8
Department of Anatomy and Cell Biology, Institute of Biomedicine, Universityof Oulu, FI-90014 Oulu, Finland

The delta opioid receptor (DOR) is one of the three opioid
receptors involved in pain and mood modulation. The human(h) DOR carries a non-synonymous single nucleotide T80G
polymorphism that replaces Phe at position 27 with Cys in thereceptor N terminus. The hDOR variants display markedly different
behavior during their “life cycle” that governs the number offunctionally active receptors at the cell surface. The variants mature
ZLWKVLPLODUVORZNLQHWLFVEXWGLIIHULQPDWXUDWLRQHIÀFLHQF\7KH
nascent receptors are subjected to N-glycan dependent quality
control that retains the precursors in the endoplasmic reticulum(ER) and eventually targets a large percentage of the Cys27 variant
precursors, but not those of the Phe27 variant, to ER–associateddegradation (ERAD). In addition, the Cys27 variant precursors
WKDWH[LWWKH(5DQGUHDFKWKHFHOOVXUIDFHDUHXQVWDEOHDQGDUH
internalized constitutively. These differences result in a lower
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membrane. The dissimilarities are increased further when the two
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VLWHDUHPXWDWHGUHVXOWLQJLQH[SUHVVLRQRIOLJDQGELQGLQJ
incompetent Cys27 variant at the cell surface. Importantly, in
co-transfected cells the Cys27 variant acts in a dominant negative
manner and impairs cell surface delivery of the Phe27 variant. It
impairs conversion of the Phe27 variant precursors to the matureform and redirects them to ERAD. This occurs via dimerization in
the ER. Nevertheless, the unpaired Cys residue in the N terminusof the Cys27 variant does not cause a major folding defect because
pharmacological characteristics of the variants are indistinguishable
and opioid antagonists, acting as pharmacological chaperones, are
DEOHWRHQKDQFHPDWXUDWLRQRIERWKYDULDQWVZKHWKHUH[SUHVVHG
individually or simultaneously. The hDOR is thus a G proteincoupled receptor that is predisposed to premature targeting to
ERAD, most likely because of its relatively slow folding rate in the
ER. It is amenable to rescue by pharmacological chaperones thatstabilize newly-synthesized receptors, and eventually enhance their(5H[SRUWDQGLQFUHDVHWKHVWHDG\VWDWHUHFHSWRUOHYHODWWKH
plasma membrane.

Ligands of G-protein coupled receptors (GPCRs) transduce signals
IURPWKHH[WHUQDOHQYLURQPHQWDFURVVWKHSODVPDPHPEUDQHWR
induce an appropriate cellular response. We have found a novel
mechanism whereby some GPCRs are able to amplify or reduce
26

International Narcotics Research Conference 2013

PROGRAM ABSTRACTS+27723,&6

0823,2,'5(&(37256+<3(532/$5,=(5(63,5$725<&21752//,1*.g//,.(5)86(1(85216
Levitt, E.S.1$EGDOD$3/23DWRQ-)52, Williams, J.T.1
Vollum Institute, Oregon Health & Science University, Portland, OR, USA
School of Physiology and Pharmacology, University of Bristol, Bristol, UK

1
2

Respiratory depression is the primary cause of death fromRSLRLGRYHUGRVH7KHSUH%RW]LQJHUFRPSOH[LQWKHPHGXOODKDV
traditionally been considered the main area responsible for opioidinduced respiratory depression. However, in the rat working heartbrainstem preparation, injection of the mu opioid agonist DAMGO(60 – 120 nl of 1 mM) into the Kölliker-Fuse (KF) area of the ponscaused disruption of the normal respiratory cycle. The primaryeffect was a loss of post-inspiratory activity, which is characteristicof silencing the KF. In whole-cell recordings from KF neurons
contained in rat brain slices, activation of mu opioid receptors
hyperpolarized KF neurons by opening G protein-coupled inwardly
rectifying potassium (GIRK) channels. The outward GIRK current
produced by [Met5]enkephalin (ME) was concentration-dependent,
reversed at the potassium equilibrium potential and was blocked
by BaCl2. In locus coeruleus neurons, a saturating concentrationof ME causes robust desensitization of the mu opioid receptorinduced current. In KF neurons, a saturating concentration of ME
caused only modest desensitization. The current produced by a
VDWXUDWLQJFRQFHQWUDWLRQRI0(GHFOLQHGE\GXULQJDWHQ
minute perfusion while the effectiveness of an EC50 concentration
RI0(ZDVUHGXFHGE\7KHFXUUHQWSURGXFHGE\DQEC50 concentration of ME recovered gradually over 30 minutes.
Pretreatment with the protein kinase C activator PDBu (100 nM)
did not alter desensitization or recovery from desensitization. Mu
opioid receptors on respiratory-controlling neurons in the KF only
modestly desensitize, which correlates with the modest tolerance
to respiratory depression observed during prolonged opioid
treatment.

02/(&8/$50(&+$1,6062)72/(5$1&(7223,2,',1:$5'5(&7,)<,1*327$66,80
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6,*1$//,1*&203/(;(67+$7$5(02'8/$7('%<E$55(67,1
*Nagi, K.1,2&KDUÀ,1,2, Hébert,T.43LQH\UR*1,2,3
Ste-Justine Hospital Research Center, 2 Dept Pharmacology and 3 Dept
Psychiatry, University of Montreal and 4 McGill University, Montreal, Quebec,
Canada1

Monitoring of G-protein coupled receptor (GPCRs) interactions
has revealed that receptors, G proteins and downstream effectors
UHDFKWKHPHPEUDQHDVDVLJQDOOLQJXQLW7KHVHVLJQDOOLQJFRPSOH[HV
maintain their integrity during early stages of receptor activation
implying that proteins responsible for receptor regulation are
PRVWSUREDEO\UHFUXLWHGWRWKHFRPSOH[UDWKHUWKDQWRLVRODWHG
receptors. Here we were interested in the regulation of DOR.LUVLJQDOOLQJFRPSOH[HV,QDÀUVWVHULHVRIH[SHULPHQWVZHXVHG
%5(7DQGFRLPPXQRSXULÀFDWLRQDVVD\VWRHVWDEOLVKLI.LU
channels constitutively associate with heterotrimeric G protein
subunits and delta opioid receptors (DORs). We then showed
WKDWDOOFRPSOH[FRPSRQHQWVUHPDLQHGDVVRFLDWHGDIWHUVXVWDLQHG
activation (30 min) of the receptor with SNC-80 or DPDPE (1µM). We further observed that DOR stimulation (SNC-80 1 µM; 30min) induced Earr2 recruitment towards DORs, GEJ subunits andwww.dcconferences.com.au/inrc2013/

Kir3.1/3.2 channels, and established that receptors and channels
H[SUHVVHGLQSULPDU\QHXURQDOFXOWXUHZHUHERWKLQWHUQDOL]HGE\
SNC-80, DPDPE and morphine. However, the internalization of.LUFKDQQHOVZDVVSHFLÀFWR'25DQGZDVQRWREVHUYHGIROORZLQJ
stimulation of GABAB receptors which also use Kir3 channels aseffectors. Moreover, receptors and Kir3 channels colocalized witheach other after stimulation with SNC-80 and morphine agonists
but only SNC-80 induced colocalization of Earr2 with DORs
and Kir3 channel subunits. Taken together, these data show that
'25VDQG.LUFKDQQHOVIRUPFRQVWLWXWLYHFRPSOH[HVWKDWUHPDLQ
associated during late stages of receptor activation and indicatethat regulatory proteins such as EDUUUHFRJQL]HWKHVHFRPSOH[HV
as a unit, causing simultaneous internalization of the receptor andthe effector.

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Simonson, B.1, Morani, A.1, Ewald, A.1, Walker, L.1,
3ULVLQ]DQR72, *Kivell, B.1
School of Biological Sciences,Victoria University of Wellington, New Zealand,
Department of Medicinal Chemistry, School of Pharmacy, University of
Kansas1
2

Activation of kappa-opioid receptors (KOPr) by Salvinorin A (SalA) attenuates cocaine prime-induced drug-seeking in rats withfewer side effects compared to traditional arylacetamide KOPr
agonists. In the present study, we investigate the behavioural
DQWLDGGLFWLRQHIIHFWVDQGVLGHHIIHFWSURÀOHRIWZRSRWHQWORQJHUDFWLQJ6DO$DQDORJXHVPHWKR[\PHWK\O6DOYLQRULQ% 0206DO
% DQGHWKR[\PHWK\OVDOYLQRULQ% (206DO% DQGHYDOXDWHWKHLU
ability to modulate the function of the dopamine transporter(DAT). Our aim is to identify novel anti-addiction compoundswith reduced side effects and identify their mechanism of action.
%RWK0206DO%DQG(206DO%VLJQLÀFDQWO\DWWHQXDWHFRFDLQH
prime-induced drug-seeking behaviour in cocaine self-administeringrats at a dose of 0.3 mg/kg. However, increased immobility anddecreased swimming in the forced swim test indicates depressive
side effects similar to those seen in the parent compound, Sal A.
MOM Sal B rapidly increases DAT function in tissue taken from
WKHUDWVWULDWXPQXFOHXVDFFXPEHQVDQGPHGLDOSUHIURQWDOFRUWH[using rotating disk electrode voltametry techniques. The dopamineXSWDNHNLQHWLFVLQWKHQXFOHXVDFFXPEHQVVKRZHGDVLJQLÀFDQWLQFUHDVHLQ9PD[  ZLWKQRFKDQJHLQ.P  FRPSDUHGWRFRQWUROV 9PD[  DQG.P 
0.6) (p<0.05)), indicating an increase in the number of functional
WUDQVSRUWHUVZLWKQRFKDQJHLQGRSDPLQHELQGLQJDIÀQLW\7KH
effects on DAT function were reversed by pre-incubation with
KOPr antagonist nor-BNI. Both MOM Sal B (p<0.01) and EOM
Sal B (p<0.05) rapidly phosphorylate ERK in vitro in a KOPrGHSHQGHQWPDQQHUXVLQJSKRVSKRVSHFLÀFDQWLERGLHVDQG:HVWHUQ
blotting techniques. Inhibition of ERK activation by U0126, a MEK
inhibitor also prevented the KOPr mediated increases in DATfunction. This study shows two novel potent Sal A analogues thathave behavioural anti-addiction effects and modulate DAT function
in a KOPr and ERK dependent manner.
Supported by Health Research Council of New Zealand, Neurological Foundationof New Zealand, and Wellington Medical Research Foundation.

27

PROGRAM ABSTRACTS

6<0326,80

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Baimel, C. and Borgland, S.L.
Department of Anesthesiology, Pharmacology and Therapeutics, University of
British Columbia,Vancouver, Canada

Dopamine neurons in the ventral tegmental area (VTA) are a
key target of addictive drugs and neuroplasticity in this region
may underlie some of the core features of addiction. All drugs
RIDEXVHLQGXFHDQ/73OLNHSRWHQWLDWLRQRIH[FLWDWRU\LQSXWVto VTA dopamine neurons. Hypocretin (hcrt), also known as
RUH[LQLVDODWHUDOK\SRWKDODPLFQHXURSHSWLGHUHOHDVHGLQWRWKH
97$WKDWH[HUWVPRGXODWRU\HIIHFWVRQDYDULHW\RIEHKDYLRUV
produced by drugs of abuse. Acute application of hcrt potentiates
H[FLWDWRU\V\QDSWLFWUDQVPLVVLRQLQWKH97$DQGLQKLELWLRQRIKFUWsignaling blocks both cocaine-induced plasticity and behavioral
sensitization. However, the role of hcrt on the plasticity inducedby other classes of abused drugs is unknown. We determined if
hypocretin action was necessary for morphine-induced synaptic
plasticity in VTA dopamine neurons using whole-cell patch clamp
electrophysiology in rat horizontal brain slices including the VTA.
Morphine potentiated glutamatergic synapses by a pre-synaptic
increase in glutamate release and by a post-synaptic change in
AMPAR number or function, likely including a switch in subunit
composition. Systemic or intra-VTA administration of SB 334867, ahcrt receptor-1 antagonist, blocked a morphine-induced increase in
the AMPAR/NMDAR ratio, morphine-induced increases in AMPARmEPSC frequency and amplitude, as well as morphine-induced$03$5UHGLVWULEXWLRQPHDVXUHGE\DFKDQJHLQUHFWLÀFDWLRQ7KHVH
results support a role for hcrt signaling in both pre-and postsynaptic potentiation of glutamatergic transmission in the VTA bymorphine. Because hcrt signaling is required for plasticity inducedby both morphine and cocaine, hcrt may function as a gate keeper
for drug induced plasticity of dopamine neurons.

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5(:$5'$1''58*6((.,1*0XVFKDPS-:3K'
Department of Psychiatry, Harvard Medical School, McLean Hospital,
Belmont, MA 02478+\SRFUHWLQ RUH[LQ DQGG\QRUSKLQDUHQHXURSHSWLGHVZLWKRSSRVLQJDFWLRQVRQPRWLYDWHGEHKDYLRU2UH[LQLVLPSOLFDWHG
LQPDLQWDLQLQJDURXVDODQGLQWKHUHZDUGLQJHIIHFWVRIIRRGVH[and drugs of abuse. Conversely, dynorphin is implicated in thedysphoric effects of stress that often accompany or precipitate
depressive-like states. We have found that, despite their opposing
DFWLRQVRQPRRGDQGEHKDYLRURUH[LQDQGG\QRUSKLQDUHSDFNDJHG
in the same synaptic vesicles within the hypothalamus, the major
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JHQHWLFGLVUXSWLRQRIRUH[LQIXQFWLRQDWWHQXDWHVWKHUHZDUGLQJ
effects of rewarding lateral hypothalamic (LH) electrical stimulation,
eliminates cocaine-induced motor impulsivity, and markedly

28

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reversed by concomitant disruption of dynorphin signaling at kappa
RSLRLGUHFHSWRUV:HDOVRGHPRQVWUDWHWKDWFRQFRPLWDQWRUH[LQ
DQGG\QRUSKLQH[HUWRSSRVLQJHIIHFWVRQLQYLWURH[FLWDELOLW\RI
ventral tegmental area (VTA) dopamine (DA) neurons, a prominent
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E\G\QRUSKLQEORFNDGH2XUÀQGLQJVLGHQWLI\DQRYHOFHOOXODU
mechanism that regulates the function of midbrain DA systems,
disruption of which could contribute to the pathophysiology ofdisorders characterized by dysregulation of motivated behavior
such as addiction and depression.

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ALCOHOL
Srinivasan, S.2, Simms, J.A.2, Hopf, F.W.2, Bonci, A.3,
*Bartlett, S.E.1
1
Translational Research Institute, Queensland University of Technology,
Brisbane, Queensland, Australia 2 Ernest Gallo Clinic and Research Center,
University of California San Francisco, CA 3 National Institute of Drug Abuse,
Maryland, USA

2UH[LQFRQWDLQLQJQHXURQVSURMHFWIURPWKHODWHUDOK\SRWKDODPXV
to the ventral tegmental area (VTA) and nucleus accumbens(NAc), two brain regions that comprise the mesolimbic ‘reward
SDWKZD\·0DQ\LQYHVWLJDWRUVKDYHVKRZQWKDWRUH[LQUHFHSWRUV2[5V LQWKH97$SOD\DUROHLQWKHPRWLYDWLRQDOGULYHIRU
addictive substances such as morphine, cocaine and alcohol.
)XUWKHUPRUHWKH2;5SOD\VDVSHFLÀFUROHLQHWKDQROVHOI
administration, cue and stress-induced relapse, with a more limited
role for OX2R being shown. We use two distinct methods tomodel ethanol and sucrose reinforcement and consumption. The
HWKDQROVHOIDGPLQLVWUDWLRQ 6$ SURWRFRODOORZVXVWRH[SORUH
the motivational salience/reinforcing properties of ethanol andthe intermittent access (IA) protocol models voluntary ‘bingelike’ ethanol consumption in the home cage. We have shown that
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and sucrose self-administration and consumption in Long-Evansrats and that intra-ventral tegmental area (VTA) infusions, but not
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not sucrose self-administration (Srinivasan et al., 2012). To begin to
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circuitry, we are using electrophysiological recordings in naive and
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2[%LQFUHDVHGÀULQJLQSUHVXPHGGRSDPLQHUJLFQHXURQVLQ97$
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that other brain regions are also involved. The results demonstrate
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ethanol and sucrose mediated behaviors.

International Narcotics Research Conference 2013

PROGRAM ABSTRACTS+27723,&6

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675(66,1'8&('&2&$,1(5(/$36(*Chiou, L.C.1, 2, 3,Tung, L.W.1, Lu, G.L.1&KDQJ/<1,Teng,
S.F.3<X/4, Hung, M.S.5, Mackie, K.6DQG=LPPHU$7.
Grad Inst Pharmacol, 2 Grad Inst Brain and Mind Sciences, 3 Dept
3KDUPDFRO&RO0HG1DWLRQDO7DLZDQ8QLYHUVLW\7DLSHL7DLZDQ4 Inst Behav
0HG&RO0HG1DWLRQDO&KHQJ.XQJ8QLYHUVLW\7DLQDQ7DLZDQ5 Inst Biotech
3KDUPDFHXW5HV1DWLRQDO+HDOWK5HVHDUFK,QVWLWXWHV=KXQDQ0LDROL7DLZDQ
6Gill Center and Dept Psychol Brain Sci, Indiana University, Bloomington,,QGLDQD86$7 Inst Mol Psychiat, University of Bonn, Bonn, Germany.
1+\SRWKDODPLFRUH[LQQHXURQVSURMHFWWRWKHYHQWUDOWHJPHQWDODUHD(VTA) whose dopaminergic activity is crucial for reward seeking.
$FWLYDWLRQRIK\SRWKDODPLFRUH[LQQHXURQVKDVEHHQUHSRUWHG
to be involved in stress-induced reward seeking. Here, we haverevealed a novel mechanism for stress-induced cocaine seeking,
LQYROYLQJRUH[LQLQGXFHGHQGRFDQQDELQRLG H&% VLJQDOLQJLQWKH
97$2UH[LQ$LQKLELWHG*$%$HUJLFWUDQVPLVVLRQLQ97$VOLFHV
via a presynaptic mechanism. This effect was antagonized by OX1(SB 334867) and CB1 (AM 251), but not OX2 (Compound 29),
antagonists, mimicked by a CB1 agonist (WIN 55,212-2) andprevented by internally applied GDP-ES and a phospholipase C
inhibitor (U73122). JZL 184, an inhibitor of monoglyceral lipasethat hydrolyses 2-arachydonoylglycerol (2-AG), an eCB, potentiated
DQGSURORQJHGWKHHIIHFWRIRUH[LQ$LQ97$VOLFHV&RQVLVWHQWO\WKHHIIHFWRIRUH[LQ$ZDVSUHYHQWHGE\WHWUDK\GUROLSVWDWLQDQ
inhibitor of diacylglycerol lipase, a 2-AG synthesizing enzyme. In a
conditioned place preference (CPP) test in mice, a 30 min-restraint
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QHXURQVLQWKHK\SRWKDODPXVVLJQLÀFDQWO\UHLQVWDWHGH[WLQJXLVKHG
cocaine CPP. This acute stress-induced cocaine reinstatement
was prevented by SB 334867 and AM 251, and was abolished in
CB1 receptor-knockout mice. These results suggest that acute
UHVWUDLQWVWUHVVDFWLYDWHVK\SRWKDODPLFRUH[LQQHXURQVWRUHOHDVH
RUH[LQVZKLFKWKHQDFWLYDWHSRVWV\QDSWLF2;UHFHSWRUVRQ
VTA dopaminergic neurons, through a Gq-protein coupledphospholipase C and diacylglycerol lipase enzymatic cascade,generating 2-AG. This eCB then retrogradely inhibits GABA releasethrough presynaptic CB1 receptors, leading to disinhibition of VTA
dopamine neurons and cocaine seeking behavior. (Supported by
the grants NSC101-2325-B002-048, NSC101-2321-B002-081 and
NHRI-EX102-10251NI).

www.dcconferences.com.au/inrc2013/

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$'0,1,675$7,21$1'(7+$12/6((.,1**Lawrence, A.J., .KRR6<Kim, A., Jupp, B., Kim, J.H., andBrown, R.M.
Florey Institute of Neuroscience & Mental Health, University of Melbourne,
VIC 3010 Australia:HZHUHWKHÀUVWWRGHPRQVWUDWHDUROHIRURUH[LQVLQERWK
ethanol consumption and cue-induced reinstatement of ethanolVHHNLQJ7KHVHHIIHFWVZHUHVSHFLÀFZLWKDGLIIHUHQWLDOHIIHFWRI
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of ethanol compared to sucrose. Moreover, we also found evidence
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DEVWLQHQFH)RVVWXGLHVVXJJHVWHGWKDWWKHSUHOLPELFFRUWH[ZDVDSRWHQWLDOORFXVZKHUHDVFHQGLQJRUH[LQHUJLFLQSXWFRXOGPRGXODWH
relapse-like behaviour. We therefore tested the role of prefrontal
cortical OX1R in cue-induced ethanol-seeking. iP rats wereWUDLQHGWRVHOIDGPLQLVWHUHWKDQRO YY DQGWKHQLPSODQWHG
with indwelling guide cannulae bilaterally targeting the prelimbic
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ZHUHVXEMHFWHGWRH[WLQFWLRQ,PPHGLDWHO\SULRUWRFXHLQGXFHG
reinstatement, rats were microinjected with either vehicle or SB334867 (3µg/side; 300nl/side). Antagonism of prelimbic OX1R with6%VLJQLÀFDQWO\DWWHQXDWHGFXHLQGXFHGUHLQVWDWHPHQWRIHWKDQROVHHNLQJLQDQDQDWRPLFDOO\VSHFLÀFPDQQHUZLWKQRHIIHFW
on sucrose-seeking. While a role for OX1R has been establishedin both ethanol reinforcement and ethanol-seeking behaviour, the
UROHRIRUH[LQUHFHSWRUV 2;5 LQWKHVHEHKDYLRXUVLVOHVVFOHDU
We therefore sought to determine the role of central OX2R inethanol-taking and ethanol-seeking behaviour. iP rats were trained
WRVHOIDGPLQLVWHUHWKDQRO ZY RUVXFURVH ZY LQ
the presence of reward-associated cues before being implantedwith indwelling guide cannulae. The selective OX2R antagonist
TCS-OX2-29 was administered intracerebroventricularly (ICV) toassess its effect on operant self-administration and cue-induced
UHLQVWDWHPHQWIROORZLQJH[WLQFWLRQ)ROORZLQJLFYLQMHFWLRQ7&6
OX2-29 reduced self-administration of ethanol, but not sucrose.
Despite reducing ethanol self-administration, TCS-OX2-29 had
no impact on cue-induced reinstatement of ethanol-seeking. To
determine where in the brain OX2R were acting to modulate
ethanol self-administration, TCS-OX2-29 was microinjected intoeither the shell or core of the nucleus accumbens (NAc). IntraNAc core, but not shell, infusions of TCS-OX2-29 decreased
responding for ethanol. Thus, OX2R in addition to OX1R may
represent a potential therapeutic target for the treatment of
alcohol use disorders. However, unlike OX1R, no impact of OX2R
antagonism was observed on cue-induced reinstatement, suggestinga more prominent role for OX2R in ethanol self-administrationcompared to cue-conditioned ethanol-seeking.

29

PROGRAM ABSTRACTS

6<0326,80

$68%6(72)63,1$/$1$/*(6,&6<1(5*<
5(48,5(6$&7,9$7,212)3527(,1.,1$6(&(36,/21
Wilcox, G.L.1,4,5, Schuster, D.J.1, Kitto, K.F.1, Metcalf, M.D.6,
Overland, A.C.1, Stone, L.S.2, Fairbanks, C.A.1,3,4
Departments of 1Neuroscience, 3Pharmaceutics, 6Medicinal Chemistry,4
Pharmacology, and 5Dermatology, University of Minnesota, Minneapolis, MN,
55455 (USA), and 2Faculty of Dentistry, Alan Edwards Center for Researchon Pain, McGill University, Montreal, Quebec (Canada).

We recently showed that spinal synergistic interactions between
delta opioid receptors (delta-ORs) and alpha2A adrenergic
receptors (alpha2A-ARs) require protein kinase C (PKC). To
identify which PKC isoforms contribute to analgesic synergy, we
HYDOXDWHGWKHHIIHFWVRIYDULRXV3.&LVRIRUPVSHFLÀFSHSWLGH
inhibitors on synergy between delta-ORs and alpha2A-ARs using
WKHWDLOÁLFNDVVD\RIWKHUPDOQRFLFHSWLRQLQPLFH2QO\D3.&
epsilon inhibitor abolished synergy between a delta-OR agonist
and an alpha2A-AR agonist. We tested a panel of combinationsof opioid and adrenergic agonists in PKC-epsilon knockout miceand found that all four combinations of a delta-OR agonist andan alpha2A-AR agonist required PKC-epsilon for antinociceptivesynergy. None of the combinations of a mu-OR agonist with analpha2-AR agonist required PKC-epsilon. However, screening
combinations of a broad range of mu- and delta-OR agonists in the
PKC-epsilon knockout mice revealed a subset that required PKCepsilon for analgesic synergy; only three delta agonists, deltorphin,DQG,,DQGR[\PRUSKLQGROHVKRZHG3.&HSVLORQGHSHQGHQW
synergy with morphine, the only mu-OR agonist to show thisGHSHQGHQFH,PPXQRKLVWRFKHPLVWU\FRQÀUPHGWKDW3.&HSVLORQ
could be found in the population of peptidergic primary afferent
nociceptors where delta-ORs and alpha2A-ARs have been foundWRH[WHQVLYHO\FRORFDOL]H,PPXQRUHDFWLYLW\IRU3.&HSVLORQZDV
found in the majority of dorsal root ganglion neurons, and intenselylabeled laminae I and II of the spinal cord dorsal horn. Protein
kinase C-epsilon is widespread in the spinal nociceptive system,
DQGLQSHSWLGHUJLFSULPDU\DIIHUHQWVLWDSSHDUVWREHVSHFLÀFDOO\involved in mediating the synergistic interaction between deltaORs and both mu-ORs and alpha2A-ARs.
Acknowledgements:The authors acknowledge support from NIH to GLW (R01DA015238) and DJS (T31-NS063634).

&$11$%,12,'23,2,',17(5$&7,216
0(&+$1,606)25,03529,1*3$,15(/,():LOVRQ3RH$
Opioids and cannabinoids interact in several clinically usefulways. Co-administration of these drugs produces acuteantinociceptive synergy, and there is a bi-directional enhancementof antinociception when one drug is administered prior to theother. Furthermore, cannabinoid administration attenuates thedevelopment of opioid tolerance. These effects are at least
partially mediated by the midbrain periaqueductal gray (PAG),
which is critically involved in descending pain modulation, opioid
tolerance, and withdrawal. Therefore, the PAG is an ideal brain
region to study the cellular and molecular mechanisms underlying
the interactions between opioids and cannabinoids. Within the
PAG, mu-opioid and CB1 receptors frequently co-localize inthe same neurons. At the presynaptic level, both mu-opioid andcannabinoid agonists inhibit GABAergic synaptic transmission in aconcentration-dependent manner. Cannabinoid inhibition of evoked
GABAergic inhibitory postsynaptic currents (IPSCs) is unaffected30

by chronic morphine treatment (CMT). Furthermore, endogenous
cannabinoid modulation of GABAergic transmission is unaffectedby CMT. However, CMT produces a rapid depression of evoked
IPSCs during high-frequency repetitive stimulation. This short-term
synaptic plasticity of synapses to presynaptic bursts is likely tohave important functional consequences within PAG, and we are
FXUUHQWO\H[DPLQLQJWKHHIIHFWRIFDQQDELQRLGVRQWKLVVKRUWWHUP
plasticity.

(;3/2,7,1*%(7:((123,2,'',))(5(1&(672(1+$1&($1$/*(6,$$1',03529(72/(5$%,/,7<Smith, M.T.
Centre for Integrated Preclinical Drug Development & School of Pharmacy,
The University of Queensland, St Lucia Campus, Brisbane, Queensland,
Australia. 4072

2[\FRGRQHDQGPRUSKLQHDUHWZRVWURQJRSLRLGDQDOJHVLFVXVHG
clinically to alleviate moderate to severe post-operative pain
as well as chronic cancer pain. Although both are members of
the benzomorphan class of opioid analgesics, they have subtle
VWUXFWXUDOGLIIHUHQFHVVXFKWKDWR[\FRGRQHKDVDPHWKR[\group, a 6-ketone, absence of the 7,8 double bond and addition
RIDK\GUR[\JURXS5HVHDUFKLQP\ODERUDWRU\PRUHWKDQ\HDUVDJRVKRZHGVLJQLÀFDQWEHWZHHQRSLRLGGLIIHUHQFHVLQ
the in vivoSKDUPDFRORJLFDOSURÀOHVRIPRUSKLQHDQGR[\FRGRQH
in rats following supraspinal bolus dose administration, a dosing
route that avoids potentially confounding pharmacokinetic effects
DVVRFLDWHGZLWKV\VWHPLFGRVLQJ7KHVHGLIIHUHQFHVZHUHFRQÀUPHG
by supraspinal pretreatment of rats with selective P-, G- andN-opioid receptor antagonists1 and by that fact that there was no
FURVVWROHUDQFHEHWZHHQVXSUDVSLQDOEROXVGRVHVRIR[\FRGRQH
in rats already rendered tolerant to intravenous morphine2.
)XUWKHUPRUHFRDGPLQLVWUDWLRQRIPRUSKLQHDQGR[\FRGRQHLQ
rats by supraspinal and systemic dosing routes produced analgesic
synergy with markedly reduced central nervous system side-effects
compared with equi-analgesic doses of either opioid alone3. Morerecently, randomized, double-blind clinical studies in patientswith moderate to severe post-operative pain administered a dual
PRUSKLQHR[\FRGRQHWUHDWPHQWPRUSKLQHDORQHRUR[\FRGRQH
alone, showed that at equi-analgesic doses, the dual morphine/
R[\FRGRQHFRPELQDWLRQWUHDWPHQWKDGDVXSHULRUDGYHUVHHYHQWSURÀOHFRPSDUHGZLWKHLWKHURSLRLGDORQH
References: 5RVV)%6PLWK077KHLQWULQVLFDQWLQRFLFHSWLYHHIIHFWVRIR[\FRGRQH
appear to be kappa opioid receptor mediated. Pain, 73: 151-57, 1997.
 1LHOVHQ&.HWDO$QWLDOORG\QLFHIÀFDF\RIWKHF-conopeptide, Xen2174,
in rats with neuropathic pain. Pain 118: 112-124, 2005.
 5RVV)%HWDO&RDGPLQLVWUDWLRQRIVXEDQDOJHVLFGRVHVRIR[\FRGRQHDQG
morphine markedly improves pain relief with reduced side-effects in rats.
Pain, 84: 421-428, 2000.
4. Blumenthal et al. Postoperative intravenous morphine consumption,
pain scores, and side-effects with perioperative oral controlled-release
R[\FRGRQHDIWHUOXPEDUGLVFHFWRP\$QHVWK$QDOJ
 5LFKDUGV3HWDO$QDOJHVLFDQGDGYHUVHHIIHFWVRIDÀ[HGUDWLRPRUSKLQH
R[\FRGRQHFRPELQDWLRQ 0R['XR LQWKHWUHDWPHQWRISRVWRSHUDWLYH
pain. J Opioid Manag 7: 217-28, 2011.
 5LFKDUGV3HWDO&RPSDULVRQRIWKHHIÀFDF\DQGVDIHW\RIGXDORSLRLGWUHDWPHQWZLWKPRUSKLQHSOXVR[\FRGRQHYHUVXVR[\FRGRQH
acetaminophen for moderate to severe acute pain after totalarthroplasty. Clin Ther 35: 498-511, 2013.

International Narcotics Research Conference 2013

PROGRAM ABSTRACTS+27723,&6

µ23,2,'5(&(37256,*1$/,1*$1'1(7,1+,%,7,21%<7$3(17$'2/,15$7/2&86
&2(58/(861(85216
Sadeghi, M., & Christie, M.J.
Pharmacology DO6, University of Sydney, NSW 2006, Australia

Opioids acting through µ-opioid receptors (MOR) are an effective
WUHDWPHQWRSWLRQIRUDFXWHSDLQVWDWHVEXWWKHVLGHHIIHFWSURÀOH
DQGOLPLWHGHIÀFDF\LQFKURQLFSDLQVWDWHVUHGXFHVLWVWKHUDSHXWLF
potential. On the other hand, inhibiting the uptake of noradrenaline
and/or activating D2-adrenoceptors seems to be effective inchronic neuropathic pain states. Tapentadol is a novel analgesic
drug that combines both moderate MOR agonism as well as
noradrenaline reuptake inhibition in a single molecule (Tzschentke
HWDO-3KDUPDFRO([S7KHU $VVXFKWDSHQWDGRO
displays analgesic activity in acute and chronic pain states and an
LPSURYHGWROHUDELOLW\SURÀOHZLWKOHVVSK\VLFDOGHSHQGHQFHOLDELOLW\than morphine.
Aim:WRLQYHVWLJDWHSRWHQF\DQGHIÀFDF\RIWDSHQWDGRODVD025
agonist and as a noradrenaline transporter (NET) inhibitor inlocus coeruleus (LC) neurons as a model system because LC cells
H[SUHVVDKRPRJHQRXVSRSXODWLRQRIQRUDGUHQHUJLFFHOOVWKDW
H[SUHVVVLPLODUOHYHOVRI025DQGD2-adrenoceptors with a smaller
contribution of D1-adrenoceptors.
Method: We performed whole-cell patch clamp recordings of
GIRK potassium channels in LC cells in horizontal brain slices of
2-5 week old rats. The effect of tapentadol as a NET inhibitor was
QRUPDOL]HGZLWKDVXSUDPD[LPDOFRQFHQWUDWLRQWKHZHOONQRZQNET inhibitor, cocaine (10µM). Results: Tapentadol is a partial
DJRQLVWDW025LQ/&FHOOVZLWK(& 0DSSUR[LPDWHO\IROGOHVVWKDQPRUSKLQHDQGDQHIÀFDF\RIUHODWLYHWR
morphine. In terms of NET inhibition, tapentadol potentiated the
HIIHFWRIH[RJHQRXVO\DSSOLHGQRUDGUHQDOLQHLQ/&FHOOVZLWKDQ
EC50 = 4 µM, which was found to be comparable to cocaine in
LC cells (EC50= 4 µM). Thus, there is potential for tapentadol as anovel analgesic drug with lower side effects due to relatively low
LQWULQVLFHIÀFDF\IRU025ZLWKDEURDGHUDQDOJHVLFSURÀOHGXHWR
a dual mode of action consisting of both MOR activation and NET
inhibition.

www.dcconferences.com.au/inrc2013/31

POSTER
LIST
'58*$'',&748$/,7<2)/,)($662&,$7(':,7+68%67,787,217+(5$3<,1,1'21(6,$*Hartati, H., 3UDVHW\R6Ismail, A., Nadjib, M. Center for Health Research University of Indonesia, Indonesia

&257,&$/02725'20,1$1&($1'029(0(175(/$7('&257,&$/327(17,$/6
$662&,$7(':,7+),1*(5)25&(352'8&7,21&KLDQJ+XDL+VLDR2IÀFHRI3K\VLFDO(GXFDWLRQ&KXQJ<XDQ&KULVWLDQ8QLYHUVLW\&KXQJ/L7DLZDQ*5.*(1(7,&32/<0253+,606$5($662&,$7(':,7+0(7+$'21(0$,17(1$1&(TREATMENT DOSE
/LX</1,5, Chung, R.H.2, Wang, S.C.16KLK<+1, Ho, I.K.1,3,4&KDQJ<61, Chen, C.H.1, Liu, C.H.1, Kuo, H.W.1, Liu, S.W.1,
Tsou, H.H.2
Center for Neuropsychiatric Research, National Health Research Institutes, Zhunan,Taiwan, 2 Division of Biostatistics and Bioinformatics, Institute ofPopulation Health Sciences, National Health Research Institutes,Taiwan, 3 Center for Drug Abuse and Addiction, China Medical University and Hospital,
Taichung,Taiwan, 4 College of Medicine, China Medical University,Taichung,Taiwan, 5 Department of Psychiatry, National Taiwan University Hospital andNational Taiwan University College of Medicine,Taipei,Taiwan
1&+$1*(6,1:,//,1*1(66726(/)0$1$*(&(175$/1(592866<67(0$021*7+(
(7+1,&*528362)&+,77$*21*+,//75$&76%$1*/$'(6+),1',1*6)520$3238/$7,21
BASED COHORT
Mollik, A.H. Md. Department of Biological Sciences, Peoples Integrated Alliance, Bogra Sadar, Bogra 5800 Bangladesh6(48(1&(2)'58*86(,1,1'21(6,$*3UDVHW\R6Subarkah, Agus Dwi Setiawan, Center for Health Research University of Indonesia, Depok City, Indonesia7+(,1)/8(1&(2)1(852/(37,&6$1'%(1=2',$6(3,17+(5$3<'58*621
7+520%2&<7(6$**5(*$7,21,19,92,13$7,(17:,7+23,2,''(3(1'(1&(Shamanskaya, M.G., Chita State Medical Academy

7+(.,1(7,&62)23,2,'5(&(37256$1'P23,2,'5(&(37256,1/,9,1*&(//6:DQJ<DQGBao, L.*, State Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Chinese Academy of Science, Shanghai, China

&52667$/.%(7:((1'(/7$23,2,'5(&(3725$1'1(59(*52:7+)$&7256,*1$/,1*
02'8/$7(61(8523527(&7,21$1'',))(5(17,$7,21,152'(17&(//02'(/6Sen, D.1<HH$1, Lammert, D.1, Huchital, M.1DQG &KHQ</1,2
12

Dept Biological Sciences, Binghamton University, the State University of New York at Binghamton, Binghamton, NY USA.
Center for Development and Behavioral Neurosciences, Binghamton University, the State University of New York at Binghamton, Binghamton, USA.

$1(:327(1723,2,'5(&(37256(/(&7,9($*21,67,163,5('%<7(75$3(37,'(6)520
$0$5,1('(5,9('3(1,&,//,80352'8&(6$1$/*(6,$:,7+287,1'8&,1*5(&(3725
(1'2&<726,6'HNDQ=16LQJK31, Jin, J.1$OHZRRG3)1, Stewart, M.13LJJRWW$01, Fontaine, F.1, Lacey, E.1, Capon, R.J.1, Sianati, S.1,
Mohammadi, S.1 & Christie, M.1
Institute for Molecular Bioscience,The University of Queensland, St Lucia, Queensland, Australia, 2Microbial Screening Technologies Pty. Ltd., Building A 28-54
3HUFLYDO5RDG6PLWKÀHOG16:$XVWUDOLD3Discipline of Pharmacology, University of Sydney, Camperdown, New South Wales, Australia

1$'(/7$23,2,'5(&(372567$7%*L*R3$7+:$</($'6721(85,7(287*52:7+$1'
1(8521$/',))(5(17,$7,21
Georganta, E., and **HRUJRXVVL=Laboratory of Cellular Signalling and Molecular Pharmacology, Institute of Biosciences and Applications,
1DWLRQDO&HQWUHIRU6FLHQWLÀF5HVHDUFK$WKHQV*UHHFH

5(3($7('0253+,1($'0,1,675$7,21'(&5($6(6$&7,9$7,212)(;75$6<1$37,&*$%$$5(&(37256,17+(3(5,$48('8&7$/*5$<$5($ 3$**Ingram, S.L.,Tonsfeldt, K.J., Suchland, K.L. & Li, M. Dept. of Neurological Surgery, Oregon Health & Science University, Portland, OR

63217$1(286*$%$5(/($6('(&5($6(6:,7+'(9(/230(17,17+(52675$/
9(17520(',$/0('8//$ 590*Li, M.,Tonsfeldt, K.J., Suchland, K.L. & Ingram, S.L., Dept. of Neurological Surgery, Oregon Health & Science University, Portland, OR32676<1$37,&%8712735(6<1$37,&0823,2,'5(&(37256,17+(35(%g7=,1*(5&203/(;'(6(16,7,=(*Lowe, J.D., Kelly, E., Henderson, G., School of Physiology and Pharmacology, University of Bristol, Bristol, UK
32

International Narcotics Research Conference 2013

POSTER LIST

678',(621'(7(50,1$7,212)%,2/2*,&$/&20326,7,212)6($:(('6&2//(&7(')5207+(681'$5%$16:,7+,1%$1*/$'(6+Mollik, A.H. Md., Department of Biological Sciences, Peoples Integrated Alliance, Bogra Sadar, Bogra 5800 Bangladesh

3+263+2,1626,7,'(.,1$6(J,17(*5$7(6&<&/,&$03$1'$.76,*1$/,1*2)7+(23,2,'5(&(3725
Schulz, S.1, Madishetti, S.2, Schneble, N.2, König, C.3, Hirsch, E.40OOHU-32, Schaible, H.G.3, Wetzker, R.2
Institute of Pharmacology and Toxicology, Jena University Hospital, Jena, Germany, 2 Institute of Molecular Cell Biology, Center for Molecular Biomedicine, Jena
University Hospital, Jena, Germany, 3 Institute of Physiology, Jena University Hospital, Jena, Germany, 4 Molecular Biotechnology Center, University of Torino,
Torino, Italy

1&255(/$7,212)(1'2&<726,6:,7+.,1(7,&62)3+263+25</$7,21$1'E$55(67,1
$662&,$7,21,123,2,'5(&(37256*Sianati, S.1, Canals, M.2, Lacey, E.3, Capon, R.J.3, $OHZRRG3)3, Christie, M.J.1, 1 Discipline of Pharmacology, University of Sydney,
NSW, 2 Faculty of Pharmacy and Pharmaceutical Science, Monash University, Parkville,VIC, 3 Institute for Molecular Bioscience, University of
Queensland, St Lucia, QLD, Australia

1$123$57,&8/$7(0253+,1(&21-8*$7(6)257+(75($70(172),1)/$00$725<3$,1*RQ]iOH]5RGUtJXH]61,a4XDGLU0$ò,a*XSWD6ò=KDQJ;òMachelska, H.1+DDJ5ò 6WHLQ&1
1
Department of Anesthesiology and Critical Care Medicine, Charité Campus Benjamin Franklin, Freie Universität Berlin, Germany. ² Institute ofChemistry and Biochemistry, Freie Universität Berlin, Germany, a contributed equally23,2,'5(&(3725&283/,1*72.&+$11(/6,13(5,3+(5$/6(1625<1(85216
5(/(9$1&()25$1$/*(6,&()),&$&<$1'63(&,(6',))(5(1&(6Nockemann, D.1, Labuz, D.1, Rouault, M.16FKPLGW<1+HSSHQVWDOO3$2 and *Stein, C.1
Dep. Anesthesiology, Charité Campus Benjamin Franklin, Freie Universität Berlin, Germany, 2Mouse Biology Unit, EMBL Monterotondo, Italy172///,.(5(&(3725&+$1*(623,2,'/,*$1'%,1',1*,1%5$,1+202*(1$7(6*Thomas, J.1, Somogyi, A.A.2, Hutchinson, M.R.3, 1, 2 Discipline of Pharmacology, University of Adelaide, Australia, 3 Discipline of Physiology,
University of Adelaide, Australia

%,$6('6,*1$//,1*2)(1'2*(128623,2,'6$77+(085(&(3725*Thompson, G., Christopoulos, A. and Canals, M. Monash Institute of Pharmacological Sciences, Drug Discovery Biology Laboratory, Parkville,
Victoria, Australia

'(9(/230(172)0,;('()),&$&<025'25/,*$1'6$1'02',),&$7,2172,03529(%/22'%5$,1%$55,(53(1(75$7,21$QDQG-31<HRPDQV/13RUWHU%DUUXV951, Jutkiewicz, E.M.2, *Traynor, J.R.2, Mosberg, H.I.
1

Department of Medicinal Chemistry, and 2 Department of Pharmacology, University of Michigan, Ann Arbor, MI, 48109, USA.

$129(/0(&+$1,60)255(675$,17675(66,1'8&('&2&$,1(5(/$36(25(;,1,1'8&('(1'2&$11$%,12,'5(752*5$'(',6,1+,%,7,21,17+(9(175$/7(*0(17$/
AREA* Tung, L.W.1, Lu, G.L.1&KDQJ/<1/HH<64<X/6, Lee, H.J.3,Teng, S.F.3, Hwang, L.L.4,5, Hung, M.S.7, Mackie, K.8,
=LPPHU$9 and Chiou, L.C.1,2,3
Grad Inst Pharmacol, 2 Grad Inst Brain and Mind Sciences, 3'HSW3KDUPDFRO&RO0HG1DWLRQDO7DLZDQ8QLYHUVLW\7DLSHL7DLZDQ4 Grad Inst Biomed Sci,
'HSW3K\VLRO7DLSHL0HGLFDO8QLYHUVLW\7DLSHL7DLZDQ6,QVW%HKDY0HG&RO0HG1DWLRQDO&KHQJ.XQJ8QLYHUVLW\7DLQDQ7DLZDQ7 Inst Biotech Pharmaceut
5HV1DWLRQDO+HDOWK5HVHDUFK,QVWLWXWHV=KXQDQ0LDROL7DLZDQ8*LOO&HQWHUDQG'HSW3V\FKRO%UDLQ6FL,QGLDQD8QLYHUVLW\%ORRPLQJWRQ,QGLDQD86$9 Inst
Mol Psychiat, University of Bonn, Bonn, Germany.

1
5

63,1$/$&7,2162)75,37$16$1'23,2,'6,1$1,1)/$00$725<3$,167$7(
Jeong, H.J., Mitchell,V. and *Vaughan, C., Pain Management Research Institute, Kolling Institute, Northern Clinical School, University of Sydney
at Royal North Shore Hospital, St Leonards, NSW 2065, Australia.

'(6,*16<17+(6,6$1'3+$50$&2/2*<2)23,2,''(/7$$*21,676:,7+$129(/)81'$0(17$/2;$=$75,&<&/2'(&$1(6758&785(:DWDQDEH<1, 2, Hayashida, K.2, Saito, D.1,Takahashi,T.1, Sakai, J.1, Nakata, E.1, Kanda,T.1, Iwai,T.2, Hirayama, S.2, Nemoto,T.2, Fujii, H.2<DPDNDZD71 and Nagase, H.2
1

Discovery Research Laboratories, Nippon Chemiphar Co. Ltd., Saitama, Japan and 2 School of Pharmacy, Kitasato University,Tokyo, Japan

7+(52/(2)',))(5(173+263+25</$7,216,7(6,15$3,''(6(16,7,=$7,212)7+(23,2,'5(&(3725<RXVXI$1, Miess, E.2, Schulz, S.2, Christie, M.J.1, 1 Discipline of Pharmacology, University of Sydney, NSW, Australia, 2 Institute ofPharmacology and Toxicology, Jena University Hospital, Friedrich Schiller University Jena, Jena Germany
www.dcconferences.com.au/inrc2013/33

POSTER LIST

'(6(16,7,=$7,212)7+(P23,2,'5(&(3725,1,17$&7$77&(//6,6&7(50,1$/
3+263+25</$7,21(66(17,$/"
Santiago, M.1'X<32, Christie, M.2, Connor, M.1, 1 The Australian School of Advance Medicine, Macquarie University1, 2109 and 2
Department of Pharmacology, University of Sydney, 2006, NSW, Australia

27. CAMKIID&21752/67+(%,2*(1(6,62)/(70,&5251$6,123,2,'72/(5$1&(*+H<DQG:DQJ=Department of Biopharmaceutical Sciences, Cancer Center, & Program for Collaborative Research in the Pharmaceutical Sciences,
University of Illinois, Chicago, 60612, USA

',65837,212)7+($&7,1&<726.(/(721'(/$<65(&29(5<2)0823,2,'5(&(3725
)520'(6(16,7,=$7,21,1/2&86&2(58/(861(85216
Arttamangkul, S.1, Schulz, S.2 and Williams, J.T.1
Vollum Institute, Oregon Health & Science University, Portland, Oregon, USA 2 Pharmacology, Jena University Hospital, Jena, Germany13+263+25</$7,212)0823,2,'5(&(37255('8&('0253+,1($1$/*(6,$,1
,1)/$00$725<3$,167$7($RNL<1, Mizoguchi, H.1, Watanabe, C.1<RQH]DZD$1, Sakurada,T.2, Sakurada, S.1
Department of Physiology and Anatomy,Tohoku Pharmaceutical University, Sendai, Miyagi, Japan 2 First Department of Pharmacology, Daiichi College of
Pharmaceutical Sciences, Fukuoka, Japan

1

7+('(/7$23,2,'5(&(3725&2175,%87(6720253+,1(72/(5$1&(,1$1$1,0$/
02'(/2)&+521,&,1)/$00$725<3$,1
Beaudry, H.1,2, Hipólito, L.2, Gupta, A.3, Devi, L.3, Gendron, L.1, Morón Concepción, J.A.2
1
Dept. of Physiology and Biophysics, Université de Sherbrooke, Sherbrooke, Canada, 2 Dept. of Anesthesiology, Columbia University Medical Center,
New York, USA, 3 Department of Pharmacology and Systems Therapeutics, Mount Sinai School of Medicine, New York, USA.

(1'272;,1(1+$1&(0(172)0253+,1(,1'8&('&21',7,21('3/$&(35()(5(1&(*Sarkar, S.10DR;1,Vigorito, M.1,2, Chang, S.L.1,3
1
Institute of Neuroimmune Pharmacology, 2 Department of Psychology, and 3 Department of Biological Sciences, Seton Hall University, 400 South Orange
Avenue, South Orange, NJ 07079, USA

$'2/(6&(17'5,1.,1*$))(&76(1'2*(128623,2,'6,1+,332&$0386$1'
$0<*'$/$,1'(3(1'(172)5($5,1*&21',7,21
3DOP6 Daoura, L., Roman, E. and Nylander, I., Neuropharmacology, Addiction & Behaviour, Department of Pharmaceutical Bioscience,
Uppsala University, Uppsala, Sweden

6,1*/(+286,1*'85,1*$'2/(6&(1&(&$86($/7(5$7,216,1%$6$//(9(/62)
(1'2*(128623,2,'3(37,'(6Roman, E., *Granholm, L., Nylander, I., Neuropharmacology, Addiction and Behaviour, Department of Pharmaceutical Bioscience, Uppsala
University, Sweden

5('8&('0,&5251$%&/867(5(;35(66,21,17+(18&/(86$&&80%(16,6,192/9(',17+(326775$16&5,37,21$/02'8/$7,2181'(51(8523$7+,&3$,1*Hamada, A.1<DPDVKLWD$16XKDUD<1, Narita, M.1,Tsuyama, J.2, Kuzumaki, N.2, Okano, H.2 and Narita, M.1
1
Dept. Pharmacol, Hoshi Univ. Sch. Med.Tokyo, Japan, 2 Dept. Physiol., Keio Univ. Sch. Med.Tokyo, Japan.

35.THE TRANSLOCATION OF THE G23,2,'5(&(372521&+2/,1(5*,&,17(51(85216,17+(
675,$780*Heath, E.1, Chieng, B.1, Christie, M.2, Balleine. B.1 1 Brain and Mind Research Institute,The University of Sydney, NSW 2050 Australia, 2Discipline of Pharmacology, University of Sydney, NSW, Australia

320&1(85216,17+(+<327+$/$08602'8/$7(3(5,3+(5$/,0081()81&7,21*Ikegami, D.17DVDNL<1, Suzuki, M.2, Aoki, K.38H]RQR<2 and Narita, M.1
Dept. Pharmacol., Hoshi Univ. Sch. Pharm. Pharmaceut. Sci., 142-8501, Japan 2 Div. Cancer Pathophysiol., NCCRI, 104-0045, Japan 3 Div. Gene and Immune
Medicine., NCCRI, 104-0045, Japan1

,192/9(0(172)'(/7$23,2,'(5*,&6<67(0,1$'8/71(852*(1(6,6*Iwasawa, C.1, Watanabe, M.1+DPDGD<1, Kuzumaki, N.1,2, Narita, M.1, Sawamoto, K.3, Narita, M.1 1 Dept. Pharmacol., Hoshi
Univ. Sch. Pham. Pharmaceut. Sci.,Tokyo, Japan 2 Dept. Physiol., Keio Univ. Sch. Med.,Tokyo, Japan 3 Dept. Dev. Regene. Biol., Nagoya City Univ., Aichi,
Japan

34

International Narcotics Research Conference 2013

POSTER LIST

',))(5(17,$/())(&762)0253+,1($1'0(7+$03+(7$0,1(217+($&7,9$7,212)
0(62/,0%,&$1'1,*52675,$7$/'23$0,1(5*,&6<67(06,ZDVH<Mori,T., Saeki,T., Iwata, N., Murata, A., Shibasaki, M., Suzuki,T. Dept Toxicology, Hoshi University School of Pharmacy andPharmaceutical Sciences,Tokyo 142-8501, Japan,192/9(0(172)6,*0$5(&(2725&+$3(521(217+((;35(66,212):,7+'5$:$/
6,*16,10253+,1('(3(1'(170,&(*Ohya, J., Mori,T., 8]DZD16XJL\DPD.6DLWRK<Shibasaki, M., Suzuki,T. Dept Toxicology, Hoshi University School of Pharmacy andPharmaceutical sciences,Tokyo 142-8501, Japan

23,2,'(5*,&0(&+$1,60,6/2&$/,=(',17+('2:1675($02)D7 NICITINIC$&(7</&+2/,1(5(&(3725%87127D4E,17+(&(175$/1(592866<67(0*Kishioka, S., Kiguchi, N., .RED\DVKL<Saika, F., Wakida, N.,<DPDPRWR&Department of Pharmacology,Wakayama Medical University
School of Medicine,Wakayama 641-0012 Japan129(/.$33$23,2,'5(&(3725$&7,9$7,1*6$/9,125,1$$1$/2*8(60206$/%$1'(206$/%$77(18$7('58*6((.,1*%(+$9,2856,17+(5$7$1'02'8/$7('23$0,1(75$163257(5)81&7,21
Simonson, B.1, Morani, A.1, Ewald, A.1, Walker, L.13ULVLQ]DQR72, *Kivell. B.1 1 School of Biological Sciences,Victoria University of
Wellington, New Zealand, 2 Department of Medicinal Chemistry, School of Pharmacy, University of Kansas

0,&52*/,$/$&7,9$7,2135(&('(6$17,23,2,'6<67(0,10253+,1($1$/*(6,&
TOLERANCE
0DWVXVKLWD<DQG8HGD+Dept of Mol Pharmacol & Neurosci, Nagasaki Univ Grad Sch of Biomed Sci, Nagasaki

())(&7,9(1(662)$0,',127$3$$*$,1670253+,1(5(6,67$171(8523$7+,&3$,1*Mizoguchi, H.1, Watanabe, C.1<RQH]DZD$1, Sakurada,T.2, Sakurada, S.1
1
2

Department of Physiology and Anatomy,Tohoku Pharmaceutical University, Sendai, Miyagi 981-8558, JapanFirst Department of Pharmacology, Daiichi College of Pharmaceutical Sciences, Fukuoka, Fukuoka 815-8511, Japan

92/817$5<$/&2+2/,17$.($1'&%$1'0235(&(3725'(16,7<,1287%5(':,67$5
RATS*Momeni, S., Bergström, L. and Roman, E., Neuropharmacology, Addiction and Behaviour, Department of Pharmaceutical Bioscience,
Uppsala University, Sweden

,192/9(0(172)7+(/21*&+$,1)$77<$&,'5(&(3725*35,1&)$,1'8&(',1)/$00$725<3$,102'(/0,&(*Nakamoto, K.1, Nishinaka,T.1, Sato, N.1, Mankura, M.2.R\DPD<3, Kasuya, F.4 and Tokuyama, S.1
1
Department of Clinical Pharmacy, Kobe Gakuin University, School of Pharmaceutical Sciences, Kobe, Japan. 2 Bizen Kasei Chemical Co. Ltd., Akaiwa
city, Okayama, Japan, 3 Laboratory of Pharmacology, Faculty of Pharmacy, Osaka Ohtani University, Osaka, Japan. 4 Biochemical Toxicology Laboratory,
Faculty of Pharmaceutical Sciences, Kobe Gakuin University, Kobe, Japan.

*3527(,1*$7(',1:$5'/<5(&7,)<,1*327$66,80 *,5. &+$11(/63/$<$35,0$5<
52/(,17+($17,12&,&(37,9(())(&72)2;<&2'21(%871270253+,1($1')(17$1</$76835$63,1$/6,7(6*Ono, H.1, Nakamura, A.1, Fujita, M.1, Shibasaki, M.2, Mori,T.2, Suzuki,T.2, Sakaguchi, G.1, Hasegawa, M.1 and Kanemasa,T.1
1
Pain & Neurology, Medicinal Research Laboratories, SHIONOGI Co., Ltd., Osaka, Japan, 2 Department of Toxicology, Hoshi University School of Pharmacy andPharmaceutical Sciences,Tokyo, Japan

+286,1*&21',7,216$))(&7%$6$/$1'$/&2+2/,1'8&('23,2,'/(9(/6,1
ADOLESCENT RATS
*3DOP6DQGNylander, I., Neuropharmacology, Addiction & Behaviour, Department of Pharmaceutical Biosciences, Uppsala University, Uppsala,Sweden7+(,17(5$&7,212)6(9(5$/$17,'(25(66$17'58*6:,7+$&87($1'&+521,&
0(7+$'21,10,&($1'3266,%/(&/,1,&$/,03/,&$7,216
Schreiber, S.1, Hostovsky, A.2, %DUDN<2, Rubovitch,V.2DQG3LFN&*2*

Department of Psychiatry,Tel Aviv Sourasky Medical Center & Tel$YLY8QLYHUVLW\6DFNOHU)DFXOW\RI0HGLFLQH Department of Anatomy,Tel-Aviv University Sackler Faculty of Medicine,Tel Aviv, Israel
127+(3(5,$48('8&7$/*5$<&2175,%87(6722;<&2'21(%871270(7+$'21($17,12&,&(37,21,17+(5$7*Reid, R.A. and Morgan, M.M., :DVKLQJWRQ6WDWH8QLYHUVLW\9DQFRXYHU:$86$www.dcconferences.com.au/inrc2013/35

POSTER LIST

,1&5($6('5(*,21$/*5$<0$77(592/80(,123,2,'5(&(3725.12&.2870,&($6'(7(50,1('%<05,92;(/%$6('0253+20(75<Sasaki, K.a,b, Sumiyoshi, A.b, Nonaka, H.b, Hall, F.S.c, 8KO*5c, Watanabe, M.d.DVDKDUD<a, Ikedae, K., Kawashima, R.b,
Soraa, I.fa
Department of Biological Psychiatry,Tohoku University Graduate School of Medicine, Sendai, Japan, bDepartment of Functional Brain Imaging, Instituteof Development, Aging and Cancer,TohokuUniversity, Sendai, Japan, cMolecular Neurobiology Branch, NIDA/IRP/NIH, Baltimore, MD, USA, dDepartment of
Anatomy, Hokkaido University Graduate School of Medicine, Sapporo, Japan, eAddictive Substance Project,Tokyo Metropolitan Institute of Medical Science,
Tokyo, Japan, fDepartment of Psychiatry, Kobe University Graduate School of Medicine, Kobe, Japan

3+<6,2/2*,&$/$1'3$7+23+<6,2/2*,&$/5(*8/$7,212),/(;35(66,21%<
12&,&(37,1253+$1,1)4 12)4 3(37,'(5(&(3725
Donica, C.L.2=KDQJ<16LPSVRQ'XUDQG&'1, *Sherry, D.M.2, 3, Gallucci, R.M.1, 2, Awwad, H.O.1, 2 and Standifer, K.M.1, 2, 3
Department Pharmaceutical Sciences, College of Pharmacy, 2Oklahoma Center for Neuroscience and 3Department Cell Biology, College of Medicine,
University of Oklahoma HSC, OKC, OK, USA
1

52. INVOLVEMENT OF P$1'G23,2,'5(&(3725)81&7,21,17+(5(:$5',1*())(&72)
 3(17$=2&,1(*Suzuki,T., Itoh,T., Saeki,T., Masukawa, D., Shibasaki, M. and Mori,T. Dept Toxicology, Hoshi University School of Pharmacy andPharmaceutical sciences,Tokyo 142-8501, Japan1$/75(;21(2175($70(172)1(8523$7+,&3$,1,10,&(/2&$//<75$16)(&7(':,7+7+(087$170823,2,'5(&(3725*(1(,163,1$/&25'
7DR3/1,3.DR-+1/DZ3<2 and Loh, H.H.2, 1*UDGXDWH,QVWLWXWHRI/LIH6FLHQFH1DWLRQDO'HIHQFH0HGLFDO&HQWHU7DLZDQ52&
Department of Pharmacology, University of Minnesota Medical School, Minneapolis, MN, USA and 3Center for Neuropsychiatric Research, National HealthResearch Institutes,Taiwan, ROC.
2

(67$%/,6+0(172)$&(175$/32676752.(3$,102'(/86,1**/2%$/&(5(%5$/
ISCHAEMIC MICE
*Tokuyama, S.1, Harada, S.1, Nakamoto, K.1, 1Department of Clinical Pharmacy, Kobe Gakuin University, School of Pharmaceutical Sciences, Kobe,Japan

(*)31230,&(Toll, L.1, Wu, J.1, Mercatelli, D.1, Kieffer, B.2 1Torrey Pines Institute for Molecular Studies, Port St. Lucie, FL, USA, 2 Institut de Génétique et de Biologie
Moléculaire et Cellulaire, Ilkirch, France

6<1(5*,60%(7:((17+('(/7$23,2,'$*21,6761&$1'$03+(7$0,1(2&&8569,$$
*/87$0$7(5*,&10'$5(&(3725'(3(1'(170(&+$1,60
Bosse, K.E.1, Jutkiewicz, E.M.1, Mabrouk, O.S.2, Schultz, K.N.2, Kennedy, R.T.2, Gnegy, M.E.1 and *Traynor, J.R.1
Department of Pharmacology and 2Department of Chemistry, University of Michigan Ann Arbor, MI, 48109, USA.

1

,192/9(0(172)6835$63,1$/$1'3(5,3+(5$/1$/2;21$=,1(,16(16,7,9(23,2,'
5(&(37256,7(6,17+((;35(66,212)P23,2,'5(&(3725$*21,67,1'8&('3+<6,&$/
'(3(1'(1&(*8]DZD1Mori,T., Sugiyama, K., 6DLWRX<Shibasaki, M., Suzuki,T. Dept Toxicology, Hoshi University School of Pharmacy andPharmaceutical sciences,Tokyo 142-8501, Japan6(/)$'0,1,675$7,212)2;<&2'21(%<$'2/(6&(17$1'$'8/70,&(',))(5(17,$//<$))(&7675,$7$/1(85275$160,77(55(&(3725*(1((;35(66,21=KDQJ<1, Schlussman, S.D.1, Butelman, E.R.1, Ho, A.1, Ott, J.2, Kreek, M.J.1
1
7KH/DERUDWRU\RIWKH%LRORJ\RI$GGLFWLYH'LVHDVHV7KH5RFNHIHOOHU8QLYHUVLW\1HZ<RUN1<,QVWLWXWHRI3V\FKRORJ\&KLQHVH$FDGHP\RI6FLHQFHV
Beijing

59. 7+(())(&72)23,2,'5(&(372532/<0253+,606215(&(3725)81&7,21*Knapman, A., Connor, M. The Australian School of Advanced Medicine, Macquarie University, Sydney, NSW, Australia

62&,$/675(66(1*$*(623,2,'02'8/$7,212)7+(/2&86&2(58/(86
125(3,1(3+5,1(6<67(0$1',1&5($6(67+(6$/,(1&(2)5(:$5'
*Chaijale, N., Curtis A.L., Snyder, K., Bhatnagar, S.,Valentino, R.J.

Childrens Hospital of Philadelphia, Philadelphia, PA. USA

0253+,1(02'8/$7(6%5($67&$1&(5&(//0(7$67$7,&327(17,$/
Imani, B.A.1, Baran, J.2&DERW3-1 3DUDW01,2 1 School of Pharmacy, University of Queensland,Woolloongabba, Australia
2

Department of Anesthesia Research, Cleveland Clinic, Cleveland, OH

36

International Narcotics Research Conference 2013

POSTERABSTRACTS
'58*$'',&748$/,7<2)/,)(
$662&,$7(':,7+68%67,787,217+(5$3<
IN INDONESIA*Hartati, H., 3UDVHW\R6Ismail, A., Nadjib, M.
Center for Health Research University of Indonesia, Depok City, Jawa Barat
Province, Indonesia

0RQGD\²SP
Rationale: Drug addict number in Indonesia is constantly high in
DURXQGWRRISRSXODWLRQLQODVWÀYH\HDUV &+58,11%
2010). Recognizably, addiction has impact on health related qualityof life (HRQol), which is very unique. To develop program for
HRQol improvement in drug users, this analysis aims to describe
its level in Indonesia.
Methods: This analysis used secondary data from 2011 survey
on drug abuse that selected sample of 2210 drug addicts drawn
by Respondent Driven Sampling method in 17 provincial capitals
in Indonesia (CHRUI-NNB, 2011). Quality of life measured was
HRQol using the generic instrument of WHO-BREF comprising offour dimensions (physic, psychology, social and environment). The
data were analyzed based on joining solely substitution therapy ornot. Presence of diseases (Hepc1, AIDS and suspected Tuberculosis)
was assessed by self report. Study ethical clearance was issued by
the University of Indonesia.
Findings: Drug users sampled averagely aged 20 to 39 years old,
PRVWO\ZHUHPDOHV  DQGZHUHPDUULHG2QO\IHZ
VXEMHFWV  ZHUHMRLQLQJVXEVWLWXWLRQWKHUDS\0HDQWRWDOVFRUH
RI+54R/RIGUXJXVHUVLQVXEVWLWXWLRQWKHUDS\VKRZKLJKHUÀJXUH
(61.7) than without this therapy (58.4), and this phenomenon isalike for all dimensions. Disease occurence may reduce HRQol,such as score for AIDS was 56.9 in contrast to non-AIDS 59.0, andscore for TB was 57.1 below 59.1 in Non-TB, yet similar score ofaround 58 for Hep C or not.
Conclusion: 'UXJDGGLFWLQYROYHGLQH[FOXVLYHO\VXEVWLWXWLRQtherapy would give positive impact to their quality of life, while theco-morbidity might reduce HRQol.

Author will be available at their posters
at the below listed date and time

2. CORTICAL MOTOR DOMINANCE AND
029(0(175(/$7('&257,&$/327(17,$/6
ASSOCIATED WITH FINGER FORCE
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2IÀFHRI3K\VLFDO(GXFDWLRQ&KXQJ<XDQ&KULVWLDQ8QLYHUVLW\&KXQJ/L
Taiwan7XHVGD\²SP
7KHÀQGLQJVRIPRYHPHQWUHODWHGFRUWLFDOSRWHQWLDOVDVVRFLDWHG
with motor effectors have shown that multiple movement
parameters were controlled by human body, such as timing, force,
orientation, and acceleration, etc. However, some constraints
VXEVLVWHGGXULQJPRYHPHQWFRQWURODQGH[HFXWLRQVXFKDVÀQJHU
LQWHUGHSHQGHQFHRUÀQJHUQHXURDQDWRPLFDOIDFWRUV,QWKHSDVWUDWHRIIRUFHGHYHORSPHQWZDVXVHGWRH[DPLQHWKHFRQWURORIÀQJHUIRUFHSURGXFWLRQ7KHPHFKDQLVPZLWKLQYDULHGUDWHVRI
force development during force production associated with higher
level of motor system still remained unknown. In terms of motor
effectors, the controls of muscles and cortical activations have
VKRZQSDUVLPRQLRXVDGDSWDWLRQLQÀQJHULQWHUGHSHQGHQFH7KH
FRQWURORIIRUFHLVWKRXJKWWREHFKDQJHGDIWHUVSHFLÀFH[HFXWLRQ
RIÀQJHUVRQERWKEHKDYLRUDODQGFRUWLFDOOHYHORIDQDO\VHV7KLVVWXG\WULHGWRDSSO\VSHFLÀFIHHGEDFNWRLQYHVWLJDWHWKHPHFKDQLVPRIQHXURPRWRUDGDSWDWLRQUHODWHGWRVL[GLIIHUHQWUDWHVRIIRUFH
development. 15 college students were required to produce
several effector-related tasks associated with force control after
motor learning. Force outputs and movement-related corticalpotentials were collected and a three-way repeated-measured
ANOVA in terms of motor learning, nominal force levels andrates of force development were used for analyses. We found that
WKHUHZHUHH[LVWLQJGLIIHUHQWFRQWUROPHFKDQLVPVZLWKLQÀQJHUVin terms of different end-effectors. The results and implications of
WKLVVWXG\ZRXOGIXUWKHUH[SORUHWKHEHKDYLRUEUDLQUHODWLRQVKLSV
,WLVVXJJHVWHGWKDWWKHXQGHUO\LQJPHFKDQLVPRIÀQJHUFRRUGLQDWLRQRUWKHFRPSOH[PRWRUV\VWHPFRXOGEHUHFRJQL]HG
neurophysiologically and cognitively.
The study was supported by National Science Council grants,Taiwan. NSC-992410-H-033-055

www.dcconferences.com.au/inrc2013/37

POSTER ABSTRACTS*5.*(1(7,&32/<0253+,606
ARE ASSOCIATED WITH METHADONE
MAINTENANCE TREATMENT DOSE
/LX</1,5, Chung, R.H.2, Wang, S.C.16KLK<+1, Ho, I.K.1,3,4,&KDQJ<61, Chen, C.H.1, Liu, C.H.1, Kuo, H.W.1, Liu, S.W.1,
Tsou, H.H.2
1
Center for Neuropsychiatric Research, National Health Research Institutes,
Zhunan,Taiwan, 2 Division of Biostatistics and Bioinformatics, Institute ofPopulation Health Sciences, National Health Research Institutes,Taiwan, 3Center for Drug Abuse and Addiction, China Medical University and Hospital,
Taichung,Taiwan, 4 College of Medicine, China Medical University,Taichung,Taiwan, 5 Department of Psychiatry, National Taiwan University Hospital andNational Taiwan University College of Medicine,Taipei,Taiwan

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Methadone maintenance treatment has been applied in clinical forheroin dependence patients. This drug has been suggested to beadministered with an initial low dose and then gradually increase
to a maintenance dose in considerance for the tolerance effectfor each individual. In this study, we tested the hypothesis thatgenetic polymorphisms in human genome may harbor susceptibility
gene for the use of methadone maintenance dose. A total of366 patients under methadone treatment were recruited withrecords of maintenance dose, plasma methadone and metabolite
FRQFHQWUDWLRQVDQGWUHDWPHQWRXWFRPHSURÀOHV7KHJHQRPLF
'1$ZHUHJHQRW\SHGXVLQJWKH$[LRP*HQRPH:LGH&+%
Array, which was population-optimized to have a better genomic
FRYHUDJHRIFRPPRQDOOHOHV 0$)! LQWKH+DQ&KLQHVHJHQRPH*HQHEDVHGDVVRFLDWLRQDQDO\VLVLGHQWLÀHGDFDQGLGDWH
gene, G protein-coupled receptor kinase 5 (GRK5), showing
JHQRPHZLGHVLJQLÀFDQFHZLWKPHWKDGRQHGRVH S [ )XUWKHUÀQHPDSSLQJZDVSHUIRUPHGWKURXJKVHOHFWLRQRI
single nucleotide polymorphisms (SNPs) within the genetic region
and in considerance the minor allele frequencies of Chinese
HWKQLFJURXSDQGWKHIXQFWLRQVZLWKLQWKHH[RQVRI*5.7KHVH
SNPs were genotyped on the same 366 patients. The SNPs
rs11819686, rs10886472, rs11198907, rs4752300, and rs1537576
of both genotype (GLM p<0.0038, FDR<0.027) and allele types(GLM p<0.0035, FDR<0.021) within the intron 1, intron 3, and
LQWURQUHJLRQVVKRZHGVLJQLÀFDQWDVVRFLDWLRQVZLWKPHWKDGRQH
maintenance dose. The allele type of rs11198898, rs10787959,
UVDQGUVDWLQWURQZHUHVLJQLÀFDQWO\DVVRFLDWHG
with the adverse reaction symptom scores of constipation (GLM,
p<0.0046, FDR<0.041). SNP rs17098759 was associated with thesymptom score of weakness (GLM, p=0.0002, FDR, p=0.0072). We
concluded that GRK5 may be an important candidate gene whichharbors biomarkers for methadone maintenance dose and adverse
reactions.

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TRACTS BANGLADESH: FINDINGS FROM A
3238/$7,21%$6('&2+257
Mollik, A.H. Md.
Department of Biological Sciences, Peoples Integrated Alliance, Bogra Sadar,
Bogra 5800 Bangladesh

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Plants form the only easily accessible healthcare alternative forthe most of ethnic population in Bangladesh. The life, tradition,and culture of ethnic groups remained almost static since last
several hundreds of years. The knowledge accrued by the ethnic
groups through generations shows the in-depth understanding

38of the forest resources. The ethnic groups’ areas were surveyed
through periodical tours in various seasons. Knowledgeable
persons of ethnic communities were contacted and information
was collected through interviews, observations, and discussions
KHOGGXULQJÀHOGVXUYH\V7KHGLVFXVVLRQVUHYHDOHGORFDOQDPHRI
plants, plant parts used by the ethnic communities. The plants
ZHUHVFLHQWLÀFDOO\LGHQWLÀHGZLWKWKHLUERWDQLFDOQDPHVDQGDXWKRU
citation. Information on 24 plants was obtained to treat central
nervous system in the observations. These plants included Aconitum
napellus L., Aegle marmelos (L.) Corrêa, Aloe vera (L.) Burm.f.,
Amomum aromaticum5R[EAndrographis paniculata (Burm.f.) Nees,
Azadirachta indica A.Juss., Bacopa monnieri (L.) Wettst., Coccinia
grandis (L.) Voigt, Cordia dichotoma G.Forst., Glycyrrhiza glabra L.,
Holarrhena pubescens Wall., Luffa cylindrica (L.) M.Roem., Nigellasativa L., Olea europaea L., Oroxylum indicum (L.) Kurz, Oxalis
corniculata L., Phyllanthus fraternus G.L.Webster, Plantago major L.,
Senna alata / 5R[ESolanum anguivi Lam., Sphaeranthus indicus
L., Terminalia bellirica *DHUWQ 5R[ETinospora cordifolia (Willd.)
Miers, and Tribulus terrestris L.. The plant materials are used singly orsometimes in combination with milk, honey, curd, water or otherplant parts. The observations support the use of plants in curing
central nervous system and noted that the formulations containa number of plants not usually used by the ethnic communities
LQRWKHUUHJLRQVRI%DQJODGHVK)XUWKHUGHWDLOHGH[SORUDWLRQDQG
collection of plants information, chemical studies, and screening for
medicinal properties will provide cost effective and reliable sourceof medicine for the welfare of humanity.

6(48(1&(2)'58*86(,1,1'21(6,$*3UDVHW\R6Subarkah, Agus Dwi Setiawan
Center for Health Research University of Indonesia, Depok City, Jawa Barat
Province, Indonesia

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Rationale: Indonesia has recognized problems related to drug
GHSHQGHQFHVKRZQE\LWVSUHYDOHQFHLQDURXQGWRRISRSXODWLRQLQODVWÀYH\HDUV &+58,11% ,GHQWLI\LQJWKH
drug type used one earlier than the other, or pattern of drug
use sequence, would help development of secondary prevention
program.
Methods: By using data collected in Indonesia surveys on drug
abuse conducted in 17 provincial capitals, there were 2207 and
2210 of drug addicts drawn through Respondent Driven Sampling
method in year 2008 and 2011 respectively. Sequence of drug use
ZDVPHDVXUHGEDVHGRQWKHVHOIUHSRUWHGDJHRIÀUVWWU\RIHDFK
drug type. Percentages of each order then were arranged, and it
leads to depict drug use sequence pattern.
Findings: Most of drug addicts were males with age ranged from13 to 58 years. The pattern of sequence of drug use shows nosubstantial difference in both survey years, with cigarette smoking
DVWKHHQWU\SRLQWDQGRYHURIWKHPFRQWLQXHGWRXVLQJJDQMD
or alcohol. Tranquilizer was preferred by around one-third. ATS
user might continue taking opiate in small probability (around 1HDUO\RQHLQÀYHDOFRKRORUEHQ]RGLD]HSLQHXVHUVWHQGHG
to forward consuming opiate. Recently ATS was more well-liked.
*HQGHULQÁXHQFHRYHUWKHSDWWHUQRIGUXJXVHVHTXHQFHZDVQRW
clear.
Conclusion: The drug use in Indonesia recorded ganja, alcoholand tranquilizer as the gate, with cigarette smoking prior to thesedrugs. Heroin remains to be consumed, hence ATS popularity
increases.

International Narcotics Research Conference 2013

POSTER ABSTRACTS

7+(,1)/8(1&(2)1(852/(37,&6$1'
%(1=2',$6(3,17+(5$3<'58*621
7+520%2&<7(6$**5(*$7,21,19,92,1
3$7,(17:,7+23,2,''(3(1'(1&(Shamanskaya, M.G.
Chita state Medical Academy

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Background: Neurotropic drugs of different groups are widely
XVHGQRZDGD\V%HVLGHVWKHFHQWUDODFWLRQWKH\LQÁXHQFHRWKHU
systems – immunity and hemostasis.
Aim:RILQÁXHQFHRIVRPHQHXURWKURSLFGUXJVRQDJJUHJDWLRQ
ability of trombocytes in vivo in patient with opioid dependence.
Methods:7KHDJJUHJDWLRQRIWKURPERF\WHVZDVH[DPLQHGE\
analyzer “Biola” (Moscow) with turbidimetric method (Born’s
method 1962). Adrenalin, ADP(10 mcg/ mL, collagen was used
as inducers. Statistical processing of results of research was
FRQGXFWHGE\XVLQJQRQSDUDPHWULFPHWKRG::LOFR[RQ0DQQ
Whitney.
Results: It was established that after injections of diazepam 1
hours - agonists of benzodiazepine receptors (0,5 mg/kg of weigh)refused the aggregation effect, in spite of adding inducers of
aggregation (adrenalin (p<0,01), collagen (p<0,01), ADP( p<0,01)). 1
hours after administration of the blocking agent D2- of dopamine
receptors of haloperidol - (0,1 mg/kg of weigh) and 1 hours afteradministration of levomepromazin - (0,5 mg/kg of weigh) also
restrained the aggregation in the setting of inducers of different
nature: adrenalin (p<0,01), ADP (p<0,01), collagen (p<0,01).
Conclusion:,QKLELWLQJLQÁXHQFHRIGLD]HSDPLVFDXVHGE\covalent bonding of drug with glycoprotein receptors GP IIb/
IIIa, which have a common RGD- consequence for bonding withÀEULQRJHQ$QRWKHUSUREDEOHPHFKDQLVPLVFRQQHFWHGZLWK
metabolism of arachidonic acid. In this case diazepam changes the
FRQIRUPDWLRQRIWKURPERF\WHPHPEUDQHZKLFKLQÁXHQFHWKH
activity of phospholipase C, phosphoinositid’s level, formation
RIWKURPER[DQH$EUDNHDFWLRQRIPRELOL]DWLRQ&DDQG
phosphorylation of protein P-47.
7KHKDORSHULGRO·VDQGOHYRPHSURPD]LQHIIHFWFDQEHH[SODLQHG
only by it’s prevention of dissociation of D to EJ- subunit ofG-protein, which leads not only to brakeage of response ofdophaminoreceptors and E-adrenoreceptors, but also others,
including D- adrenoreceptors. Just only dissociation of subunits ofG-protein can be the necessary condition of receptors’ activation.
As approval of this statement can be the fact of inhibiting of
aggregation ability in case of adding not only adrenalin, but adding
ADP and collagen, biological effects of which are connectedwith other receptors and occur in case of dissociation of D to
EJVXEXQLWRI*SURWHLQ,IWKLVVWDWHPHQWLVWUXHZHDPSOLÀHGsignalling mechanisms in blood platelet in case of agonists
adrenoreceptors actions.

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P23,2,'5(&(37256,1/,9,1*&(//6:DQJ<DQGBao, L.*
State Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology,
Chinese Academy of Science, Shanghai 200031, China

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The µ-opioid receptors (MORs) and P-opioid receptors (DORs)
are both members of the G protein-coupled receptor family. They
play important roles in the regulation of pain. The internalized
MORs are processed for recycling back to the cytoplasmic
membrane following stimulation with the selective MOR agonist
DAMGO. The internalized DORs are entered to degradation
pathway in the present of its selective agonist Deltorphin I.
www.dcconferences.com.au/inrc2013/

Previous study reports that DORs form hetero-oligomers with
MORs and DOR agonist could induce co-internalization andco-degradation of two receptors. Therefore, the behavior of thereceptor hetero-oligomers are different from MORs and DORs
H[LVWHGDORQH+RZHYHUOLWWOHLVNQRZQDERXWWKHNLQHWLFVDQG
interaction of these receptors in living cells. In the present study,
ZHXVHWRWDOLQWHUQDOUHÁHFWLRQÁXRUHVFHQFH 7,5) PLFURVFRSH
to observe the dynamics of DORs and MORs on the cytoplasmic
membrane in living cells. We have successfully constructed alive-cell imaging system for MORs and DORs, and calculated the
kinetics of these receptors in different conditions. In the presenceof their selective agonist, the movement of the receptors were
VORZHGGRZQLQDWLPHGHSHQGHQWPDQQHU7KHGLIIXVLRQFRHIÀFLHQW
of DORs was decreased more rapidly than that of MORs. Using
confocal microscopy, we further found that the internalized MORs
frequently appeared in the tubular endosomes, but DORs weremore inclined to present in vesicular endosomes. Our research
provides a approach to study the kinetics feature of the receptors
on the membrane and help us to understand the interaction of
opioid receptors.

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6,*1$/,1*02'8/$7(61(8523527(&7,21
AND DIFFERENTIATION IN RODENT CELL
MODELS
Sen, D.1<HH$1, Lammert, D.1, Huchital, M.1, and *Chen,</1,2Dept Biological Sciences, Binghamton University, the State University of
New York at Binghamton, Binghamton, NY USA.
2
Center for Development and Behavioral Neurosciences, Binghamton
University, the State University of New York at Binghamton, Binghamton, USA.
1

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Both opioid signaling and neurotrophic factor signaling have played
an important role in neuroprotection and differentiation in the
nervous system. Little is known about whether these two signaling
pathways crosstalk to each other to affect neuroprotection anddifferentiation. Previously, we found that nerve growth factor1*) FRXOGLQGXFHH[SUHVVLRQRIWKHGHOWDRSLRLGUHFHSWRUJHQH
(Oprd1, dor) mainly through PI3K/Akt/NF-kB signaling in PC12h
cells. In this study, using two NGF-responsive rodent cell model
systems, PC12h cells and F11 cells, we found the delta opioid
neuropeptide [D-Ala2, D-Leu5]enkephalin (DADLE)-mediatedneuroprotective effect could be blocked by pharmacologicalreagents, the delta opioid antagonist naltrindole, PI3 kinase
LQKLELWRU/<DQG0$3.LQKLELWRU3'7UNNLQDVH
inhibitor K252a, respectively. Western blot analysis revealed that
DADLE activated both the PI3K/Akt and MAPK pathways in the
WZRFHOOOLQHVVL51$2SUGJHQHNQRFNGRZQH[SHULPHQWVKRZHG
that the upregulation of NGF mRNA level was inhibited withconcomitant inhibition of the survival effects of DADLE in bothcell models. siRNA Oprd1 gene knockdown also attenuated the
DADLE-mediated neurite outgrowth in PC12h cells as well as
phosphorylation of MAPK and Akt in PC12h and F11, respectively.
These data together strongly suggest that delta opioid peptide
DADLE acts through the NGF-induced functional G proteincoupled delta opioid receptor to provide its neuroprotective anddifferentiating effects at least in part by regulating survival anddifferentiating MAPK and PI3K/Akt signaling pathways in rodent
neuronal cells.

39

POSTER ABSTRACTS$1(:327(1723,2,'5(&(3725
6(/(&7,9($*21,67,163,5('%<
7(75$3(37,'(6)520$0$5,1('(5,9('
3(1,&,//,80352'8&(6$1$/*(6,$:,7+287,1'8&,1*5(&(3725
(1'2&<726,6'HNDQ=16LQJK31, Jin, J.1$OHZRRG3)1, Stewart, M.1,
3LJJRWW$01, Fontaine, F.1, Lacey, E.1, Capon, R.J.1, Sianati,
S.1, Mohammadi, S.1 & Christie, M.1
1
Institute for Molecular Bioscience,The University of Queensland, St Lucia,
Queensland, Australia, 2Microbial Screening Technologies Pty. Ltd., Building$3HUFLYDO5RDG6PLWKÀHOG16:$XVWUDOLD3Discipline of
Pharmacology, University of Sydney, Camperdown, New South Wales,
Australia

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The therapeutic potential of opioid receptor agonists is often
limited by their high propensity to cause tolerance after chronic
administration. One of the possible mechanisms responsible forthe development of tolerance is the agonist-dependent modulationof receptor internalization. Based on our discovery of a D-aminoacid rich tetrapeptide from Penicillium bilaii, we have developed
a structurally novel µ-opioid receptor selective (binding Ki = 1.5
nM (µ), 300 nM (G) and 1000 nM (N)) agonist (Bilaid C2) that fails
to produce receptor internalization in contrast to other peptidic
agonists endomorphins 1 & 2, Met-enkephalin and DAMGO.
Bilaid C2 produced analgesia comparable to that of morphinewhen administered intrathecally (3 µg) but failed to produce
antinociception subcutaneously (s.c.) at doses as high as 100 mg/
kg, indicating poor blood-brain-barrier (BBB) penetration. With
the aim of improving BBB permeability, we have made further
FKHPLFDOPRGLÀFDWLRQVWR%LODLG&LQFOXGLQJWKHLQWURGXFWLRQRID
saccharide, a helical penetrating sequence, as well as enzymatically
and/or chemically reversible moieties to mask the N-terminus.
A number of these analogues produced analgesic effects throughs.c. administration at doses comparable to that of morphine.
These compounds are potential candidates for the developmentof s.c. injectable analgesics that may have altered tolerance anddependence inducing properties.

$'(/7$23,2,'5(&(372567$7%*L*R
3$7+:$</($'6721(85,7(287*52:7+
$1'1(8521$/',))(5(17,$7,21
Georganta, E., and **HRUJRXVVL=
Laboratory of Cellular Signalling and Molecular Pharmacology, Institute of%LRVFLHQFHVDQG$SSOLFDWLRQV1DWLRQDO&HQWUHIRU6FLHQWLÀF5HVHDUFK$WKHQVGreece7XHVGD\²SP
Opioid receptors are coupled to Gi/Go proteins and recentÀQGLQJVKDYHVKRZQWKDWFDQIRUPDPXOWLFRPSRQHQWVLJQDOOLQJFRPSOH[FRQVLVWLQJRIPHPEHUVRI*SURWHLQDQGWKH6LJQDO
Transducer and Activator of Transcription STAT5B. We thus
wondered whether activation of the opioid receptors could
direct differentiation and neurite outgrowth through a molecularpathway involving STAT5B and other signaling intermediates. We
demonstrate that prolonged G-opioid receptor (G-OR) activation
with opioid agonists induces STAT5B phosphorylation in Neuro2A cells. Moreover, DSLET-activation of G-OR triggers neurite
outgrowth and neuronal survival; these effects are blocked by40

SHUWXVVLVWR[LQWUHDWPHQWDQGDIWHUH[SUHVVLRQRIDGRPLQDQW
negative mutant of STAT5B (DN-STAT5B), suggesting that thesignalling pathway participating in this mechanism involves Gi/oproteins and p-STAT5B. Additional studies have shown that theZKLOH'6/(7H[SRVXUHRIQHXUREODVWRPDFHOOVLQGXFHVDPDUNHG
LQFUHDVHRIVSHFLÀFGLIIHUHQWLDWLRQPDUNHUSURWHLQVRYHUH[SUHVVLRQRIWKH'167$7%DWWHQXDWHGVLJQLÀFDQWO\WKHVHHIIHFWV7DNHQWRJHWKHURXUÀQGLQJVGHPRQVWUDWHWKDWG-OR activation leads toa number of neurotropic events via a GDi/o-linked and STAT5Bdependent manner.
This work was supported by the GSRT and the EU grant “Normolife” (LSHCCT2006-037733).

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'(&5($6(6$&7,9$7,212)(;75$6<1$37,&*$%$$5(&(37256,17+(3(5,$48('8&7$/
*5$<$5($ 3$**Ingram, S.L.,Tonsfeldt, K.J., Suchland, K.L. & Li, M.
Dept. of Neurological Surgery, Oregon Health & Science University, Portland,
OR

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Tonic inhibition of PAG output neurons by GABA is important for
the normal control of descending pain regulation. Opioids inhibit
GABA release in the PAG resulting in activation of PAG output
neurons and the descending antinociception pathway that inhibits
QRFLFHSWLYHLPSXOVHVLQWKHVSLQDOFRUG([WUDV\QDSWLF*$%$$
receptors are known to regulate the gain of inhibitory systems
DQGÀULQJWKUHVKROGVRIQHXURQV+RZHYHULWLVQRWNQRZQLIWRQLF*$%$UHOHDVHLQWKH3$*DFWLYDWHVH[WUDV\QDSWLF*$%$$
receptors or if these receptors are modulated following repeatedadministration of morphine. Male Sprague-Dawley rats werepretreated twice a day with saline or morphine (5 mg/kg, s.c.) for
3 days. This pretreatment caused a shift to the right for DAMGO
inhibition of miniature GABAergic inhibitory postsynaptic currents(mIPSCs). Superfusion of the GABAA antagonist, bicuculline (10µM), inhibited mIPSCs and blocked a resting current indicating the
SUHVHQFHRIERWKV\QDSWLFDQGH[WUDV\QDSWLF*$%$$PHGLDWHG
currents in the PAG. Bicuculline reduced the holding current by
18 ± 4 pA (n = 10) in saline pretreated rats but this effect was
VLJQLÀFDQWO\UHGXFHGLQUDWVSUHWUHDWHGZLWKPRUSKLQH S$
n = 6, t(14) = 3.46, p < 0.05). The bicuculline-induced currents
UHYHUVHGDWWKHH[SHFWHGFKORULGHHTXLOLEULXPSRWHQWLDOLQFKORULGH
(-24 ± 3 mV) and gluconate-substituted (-84 ± 2 mV) intracellularsolutions suggesting that GABAA receptors elicit a tonic current
that is inhibited by bicuculline. Tonic currents are often mediated
E\H[WUDV\QDSWLF*$%$$UHFHSWRUVFRQWDLQLQJWKHGHOWDVXEXQLWThe GABAA agonist THIP selectively activates GABAA receptors
containing delta subunits. THIP (10 µM) increased the BICblocked currents by 5-fold (84 ± 22 pA, n= 8) in opioid-naïve
slices and morphine treated slices (77 ± 24 pA, n = 6). Western
EORWDQDO\VHVIRU*$%$$GHOWDVXEXQLWVGLGQRWVKRZDVLJQLÀFDQW
difference between total GABAA-delta subunits in the PAG of
saline and morphine-pretreated rats. These results suggest thatGABAA-delta receptors are not internalized but that activation of
H[WUDV\QDSWLF*$%$$UHFHSWRUVLVGHFUHDVHGLQPRUSKLQHWROHUDQW
rats. Supported by NIH R01DA027625 (SLI).

International Narcotics Research Conference 2013

POSTER ABSTRACTS63217$1(286*$%$5(/($6('(&5($6(6:,7+'(9(/230(17,17+(52675$/
9(17520(',$/0('8//$ 590*Li, M.,Tonsfeldt, K.J., Suchland, K.L. & Ingram, S.L.
Dept. of Neurological Surgery, Oregon Health & Science University, Portland,
OR

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Recent studies have shown evidence that electrical stimulation of
the RVM produces pain facilitation in young animals but biphasic
pain facilitation and inhibition in adults. This developmental change
appears to be dependent on endogenous opioid activity prior topostnatal day 21. Opioid inhibition of GABA release onto RVM
neurons output neurons to the spinal cord is thought to be themechanism of opioid-induced antinociception in the RVM but
cellular studies have primarily studied opioid responses in young
RVM slices (< 18 days postnatal). In this study, basal spontaneous*$%$UHOHDVHLQWKHSUHVHQFHRIWHWURGRWR[LQ 77;0 ZDV
compared in young (0.07 ± 0.03 Hz; N = 5; postnatal days 8-18)
and adult rats (>35 days; N = 3; 0.03 ± 0.03 Hz, Mann-Whitney U(6, 30), p < 0.05). Interestingly, removal of TTX had little effecton spontaneous GABA release in adult rat slices (0.08 ± 0.05 Hz;
1  EXWVLJQLÀFDQWO\LQFUHDVHGVSRQWDQHRXV*$%$V,36&VLQ
young slices (0.4 ± 0.1 Hz, N = 3; Mann-Whitney U (21, 24), p < 7KLVUHVXOWLQGLFDWHVWKDWVSRQWDQHRXVÀULQJRI*$%$HUJLF
neurons intrinsic to the RVM may change during development.
Replacing the KCl-based intracellular pipette solution with CsCl
VLJQLÀFDQWO\HQKDQFHGWKHUDWHLQ\RXQJVOLFHV +]1  
2SLRLGLQKLELWLRQRIHYRNHG*$%$HUJLF,36&VZDVQRWVLJQLÀFDQWO\different with age (met-enkephalin (ME 10 µM) inhibition: young,1 FRPSDUHGWRDGXOW1  7KHDELOLW\
RI0(WRLQKLELWP,36&VZDVDOVRVLPLODULQ\RXQJ 1  FRPSDUHGWRDGXOWVOLFHV 1  +RZHYHUWKHUHZDV
a trend toward larger ME (10 µM) activation of GIRK currents inadults (52 ± 15 pA, N = 6) compared to young rats (26 ± 12 pA, N )XUWKHUH[SHULPHQWVZLOOGHWHUPLQHZKHWKHUWKHGHFUHDVHG
GABAergic tone in adult rats contributes to the observed
developmental differences in behavioral responses to electricalstimulation of the RVM. Supported by NIH R01DA027625 (SLI).

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0823,2,'5(&(37256,17+(35(%g7=,1*(5&203/(;'(6(16,7,=(*Lowe, J.D., Kelly, E., Henderson, G.
School of Physiology and Pharmacology, University of Bristol, Bristol, UK
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7KHPRVWVLJQLÀFDQWDGYHUVHFRQVHTXHQFHRIWKHXVHRIRSLDWHVLV
respiratory depression which can lead to death. Opioid agonists
suppress respiration by activating mu-opioid receptors (MOPr)
LQWKHYHQWUDOUHVSLUDWRU\FROXPQDQGPRUHVSHFLÀFDOO\ZLWKLQJOXWDPDWHUJLFQHXURQVRIWKHSUH%|W]LQJHUFRPSOH[ SUH%|W& 
which is believed to be the dominant rhythm generator. We
sought to determine if MOPrs within the preBötC undergoagonist-dependent desensitization using whole-cell patch-clamp
electrophysiology in mouse brain slices. The preBötC contains
DKHWHURJHQRXVSRSXODWLRQRIQHXURQVEXWFHOOVFRH[SUHVVLQJ
MOPrs and neurokinin-1 receptors (NK1) are thought toJHQHUDWHWKHLQVSLUDWRU\UK\WKPRIQHXURQVUHVSRQGHGWR
the MOPr agonist Met-Enkephalin (30PM) which was measured
either as an outward current in cells voltage clamped to -60mV(33.5pA±4.4pA) or as a hyperpolarization in cells current clamped
to a membrane potential of -45mV (8.2mV±2.8mV). Of the Met-

www.dcconferences.com.au/inrc2013/(QNHSKDOLQUHVSRQGLQJFHOOVVKRZHGDQLQZDUGFXUUHQWin response to the NK1 agonist [Sar9,Met(O2)11]-Substance P(500nM). We also measured GABA-B receptor responses as thesereceptors couple to the inwardly rectifying potassium channels that
presumably underlie the MOPr response. The GABA-B agonist
baclofen (20P0 SURGXFHGDQRXWZDUGFXUUHQWLQRIWKH
Met-Enkephalin responding cells. Although the MOPr currents
ZHUHVPDOOZHZHUHDEOHWRPHDVXUHVLJQLÀFDQWGHVHQVLWL]DWLRQ DIWHUPLQXWHVRI0HW(QNHSKDOLQDSSOLFDWLRQ
Because the postsynaptic Met-Enkephalin responses were small
DQGRQO\RFFXUUHGLQDVXEVHWRISUH%|W&QHXURQVZHH[DPLQHG
if presynaptic MOPrs within the preBötC might produce a morerobust inhibition of glutamatergic release since preBötC neurons
synapse onto neurons in both ipsi- and contralateral preBötC.
Met-Enkephalin (30P0 SURGXFHGDLQKLELWLRQRIWKH
DPSOLWXGHRIHYRNHGH[FLWDWRU\SRVWV\QDSWLFFXUUHQWVKRZHYHU
this inhibition remained stable over a 10 minute application.
Thus, postsynaptic MOPrs in the preBötC undergo agonistGHSHQGHQWGHVHQVLWL]DWLRQLQWKHVXEVHWRIFHOOVWKDWH[SUHVV
them, whereas presynaptic MOPrs may produce a more robust
and homogenous response that does not undergo rapid agonistLQGXFHGGHVHQVLWL]DWLRQ)XUWKHUVWXGLHVZLOOH[DPLQHZKHWKHU
tolerance develops to the MOPr effects in the preBötC after invivo treatment with opiates. (Funded by MRC)

678',(621'(7(50,1$7,212)
%,2/2*,&$/&20326,7,212)6($:(('6&2//(&7(')5207+(681'$5%$16:,7+,1
BANGLADESH
Mollik, A.H. Md.
Department of Biological Sciences, Peoples Integrated Alliance, Bogra Sadar,
Bogra 5800 Bangladesh

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The Sundarbans is the largest single block of tidal halophytic
mangrove forest in the world. The Sundarbans is a UNESCO
world heritage site covering parts of Bangladesh and Indian stateof West Bengal. Two-third of the Sundarbans is in Bangladesh
and the remaining third in West Bengal of India. Three selectedsea-weeds namely Laminaria spp., Fucus spp., and Sargassum spp.
were collected during October 2012 to March 2013 from the
6XQGDUEDQVZLWKLQ%DQJODGHVKLQRUGHUWRÀQGRXWWKHLUV\VWHPDWLF
status, and chemical & biochemical composition. High abundanceof Laminaria spp., and Fucus spp. were found in January, February,
and March 2013 respectively during the period of investigations.
7KHPD[LPXPDQGWKHPLQLPXPSHUFHQWDJHRISURWHLQZHUH
noted to be 61.21 in Laminaria spp. in the month of October 2012,
and 2.75 in Laminaria spp. in March 2013 where as the lowest
percentage of carbohydrate was observed as 11.25 in Laminaria
VSSLQ2FWREHU$QRWKHUVLJQLÀFDQWFRQVWLWXHQWLQVHDZHHGVIRUH[DPSOHDVKZDVUHFRUGHGDVLQ)XFXVVSSLQ0DUFK
DQGLQ/DPLQDULDVSSLQWKHPRQWKRI2FWREHUDVhighest, and lowest percentage respectively. The mean percentage
of highest contents of Calcium (Ca) recorded in Sargassum spp.
was 4.11±0.25 in March 2013, and the lowest in Laminaria spp.
was 1.49±0.27 in February 2013. The height Iodine (I) contents
observed in Fucus spp. was 0.113±0.025 in February 2013, and thelowest in Sargassum spp. was 0.015±0.005 in February 2013, andthe minimum contents were 175.25 ppm in Laminaria spp. January
2013. These investigations were conducted to have a nutritional
data for Laminaria spp., Fucus spp., and Sargassum spp. in order topopularize their consumption, and utilization in Bangladesh.

41

POSTER ABSTRACTS

3+263+2,1626,7,'(.,1$6(J,17(*5$7(6&<&/,&$03$1'$.7
6,*1$/,1*2)7+(23,2,'5(&(3725
Schulz, S.1, Madishetti, S.2, Schneble, N.2, König, C.3, Hirsch,
E.40OOHU-32, Schaible, H.G.3, Wetzker, R.2

&255(/$7,212)(1'2&<726,6:,7+.,1(7,&62)3+263+25</$7,21$1'
E$55(67,1$662&,$7,21,123,2,'
5(&(37256

Institute of Pharmacology and Toxicology, Jena University Hospital, Jena,
Germany, 2 Institute of Molecular Cell Biology, Center for Molecular
Biomedicine, Jena University Hospital, Jena, Germany, 3 Institute of Physiology,
Jena University Hospital, Jena, Germany, 4 Molecular Biotechnology Center,
University of Torino,Torino, Italy*Sianati, S.1, Canals, M.2, Lacey, E.3, Capon, R.J.3, Alewood,3)3, Christie, M.J.1
1
Discipline of Pharmacology, University of Sydney, NSW, 2 Faculty of
Pharmacy and Pharmaceutical Science, Monash University, Parkville,VIC,
3
Institute for Molecular Bioscience, University of Queensland, St Lucia,
QLD, Australia

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7XHVGD\²SP

1

The µ -opioid receptor (MOR) has been characterized as main
mediator of opioid signaling in neuronal cells. Gi subunits ofheterotrimeric G proteins and their inhibitory effects on cAMP
were shown to convey pain depressant signaling effects of MOR.
Recent investigations revealed PI3KJ and Akt as additional
elements of MOR signaling. PI3KJLVPDLQO\H[SUHVVHGLQFHOOV
of the immune system. Only recent investigations describe an
H[SUHVVLRQLQWKHQHXURQDOV\VWHP7KHSUHVHQWVWXG\UHYHDOV3,.J
as a connecting point of pronociceptive cAMP- and antinociceptive
PI3K/Akt –signaling pathways in neuronal cells.
The human neuroblastoma cell line SK-N-LO and dorsal root
ganglia from mice have been used to characterize MOR signaling
reactions. In both cellular systems cAMP level was manipulated
by inhibition of cAMP phosphodiesterases and stimulation of
adenylate cyclases and corresponding effects on MOR dependent
PI3K/Akt signaling have been analyzed. In order to investigate thespecial role of PI3KJLQ025VLJQDOLQJ6.1/2FHOOVH[SUHVVLRQRI
PI3KJ was stably down regulated using shRNA approach.
Stimulation of SK-N-LO cells with morphine resulted in
activation of Akt and Erk1/2. PI3KJ down–regulated cells showed
DVLJQLÀFDQWO\UHGXFHG$NWSKRVSKRU\ODWLRQ0RUHRYHUIRUVNROLQ
pretreatment of the cells resulted in inhibition of morphine
induced Akt activation. Inhibitory effects of cAMP and protein
kinase A on MOR induced stimulation of PI3KJ and Akt suggest a
conjunction of both signaling pathways.
Together our data disclose PI3KJ as a mediator of the inhibitory
action of cAMP and PKA on Akt implicating this PI3K species
as a knot point of pronociceptive cAMP/PKA signaling andantinociceptive signaling reactions of PI3K/Akt in neuronal cells.
Keywords: µ -opioid receptor, PI3KJ, cAMP, dorsal root ganglia, pain,
nocicpetion

42

Attempts to develop opioid analgesics with reduced ability to
SURGXFHWROHUDQFHDQGGHSHQGHQFHDIWHUFKURQLFH[SRVXUHKDYH
recently focused on ligand-direct signalling or biased agonism,
whereby different agonists at a G protein-coupled receptor(GPCR) can activate distinct downstream signaling pathways. We
KDYHXVHG$W7FHOOVVWDEO\H[SUHVVLQJ)/$*WDJJHG025WR
compare the ability of a number of µ-opioid receptor ligands topromote phosphorylation, Earrestin-2 association and endocytosis.
We selected a range of agonists including Endomorphin2, morphine
DQG%LODLG&ZKLFKKDYHPRGHUDWH*SURWHLQFRXSOLQJHIÀFDF\EXWZLWKDGLIIHUHQWSURÀOHLQSURGXFLQJHQGRF\WRVLVIURPUREXVWto very low respectively; and Met-enkephalin or DAMGOZLWKKLJK*SURWHLQFRXSOLQJHIÀFDF\DQGHQGRF\WRVLVFDSDFLW\
The results show a good correlation between agonist-induced
phosphorylation of Ser375 and internalization. After 3 min agonist
H[SRVXUH6HUSKRVSKRU\ODWLRQZDVPD[LPDOIRU'$0*2
Met-enkephalin and endomorphin-2, modest for morphine andvery weak for Bilaid-C2. In contrast, we observed a different
pattern in dephosphorylation rate of Ser375 between theseagonists. The fastest rate of dephosphorylation was observedwith non-internalizing agonist Bilaid-C2. The rank order of Ser375
dephosphorylation rate was Bilaid-C2 > morphine > MetHQNHSKDOLQ'$0*2!!(QGRPRUSKLQ:HXVHG%5(7DVVD\
to assess MOR-Earrestin2 interaction after agonist application. The
data indicate that agonist-induced Earrestin2 association correlates
well with Ser375 phosphorylation. DAMGO, Met-enkephalin andEndomorphin-2 produced rapid increases in BRET ratio, whereasmorphine produced smaller increase in the BRET ratio andBilaid-C2 induced very little Earrestin2 recruitment. Interestingly,
the rank order of rate of Ser375 dephosphorylation was the sameas Earrestin2 dissociation. These results demonstrate that agonists
such as endomorphin-2 with high bias to produce internalization
display relatively a slow rate of dephosphorylation and arrestin
dissociation.

International Narcotics Research Conference 2013

POSTER ABSTRACTS1$123$57,&8/$7(0253+,1(&21-8*$7(6)257+(75($70(172)
,1)/$00$725<3$,1*RQ]iOH]5RGUtJXH]61,a4XDGLU0$ò,a*XSWD6ò=KDQJ;ò0DFKHOVND+1+DDJ5ò 6WHLQ&1
1
Department of Anesthesiology and Critical Care Medicine, CharitéCampus Benjamin Franklin, Freie Universität Berlin, Germany. ² Instituteof Chemistry and Biochemistry, Freie Universität Berlin, Germany, acontributed equally
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component of many painful syndromes, including arthritis,ÀEURP\DOJLDHQGRPHWULRVLVF\VWLWLVFDQFHUWUDXPDZRXQGVand postoperative pain. Conventional opioid analgesics arelimited by side effects (e.g. sedation, apnoea, addiction, tolerance,
FRQVWLSDWLRQ 7KHLQKLELWLRQRILQÁDPPDWRU\SDLQFDQEHDFKLHYHG
by activating opioid receptors on peripheral sensory nerve
terminals. Nanocarriers have been developed to selectively activate
those receptors and prevent the distribution of opioids to healthy
QRQLQÁDPHG WLVVXHVVXFKDVEUDLQRUHQWHULFQHUYHSOH[XV+HUHZHH[DPLQHDQHZQDQRSDUWLFXODWHS+VHQVLWLYHPRUSKLQH
polyglycerol conjugate (PGM) in comparison to conventional
morphine sulphate (MS). We hypothesized that morphine is
UHOHDVHGRQO\DWDFLGLFS+LQDUDWPRGHORILQÁDPPDWRU\SDLQ
Following intravenous (i.v.) or intraplantar (i.pl.) injection into theLQÁDPHGSDZDQDQDOJHVLFHIIHFW DVPHDVXUHGE\HOHYDWHGSDZ
SUHVVXUHWKUHVKROGV337 ZDVRQO\GHWHFWHGLQWKHLQÁDPHGSDZ
and was completely abolished by i.pl. injection of the quaternary
RSLRLGDQWDJRQLVWQDOR[RQHPHWKLRGLGH 1/;0 7KHKLJKHVWGRVH
RILSO06LQFUHDVHG337LQWKHFRQWUDODWHUDOQRQLQÁDPHGSDZ
indicating central effects. Moreover, i.v. MS increased PPT dosedependently in both hindpaws and was not completely reversible
by i.pl. NLXM, suggesting the involvement of central opioid
UHFHSWRUV0LFURGLDO\VLVH[SHULPHQWVUHYHDOHGWKHSUHVHQFHRIIUHH
PRUSKLQHLQLQÁDPHGSDZVDIWHULSOLQMHFWLRQRI06RU3*06PDOO
amounts of free morphine were also detectable in the contralateral
QRQLQÁDPHGSDZKDIWHULSO067KHVHUHVXOWVVXJJHVWWKDWVXFKnew “controlled release” formulations of potent conventional
opioid analgesics could improve pharmacological therapies avoiding
central effects.

23,2,'5(&(3725&283/,1*72.
&+$11(/6,13(5,3+(5$/6(1625<
1(852165(/(9$1&()25$1$/*(6,&()),&$&<$1'63(&,(6',))(5(1&(6Nockemann, D.1, Labuz, D.1, Rouault, M.16FKPLGW<1,+HSSHQVWDOO3$2 and *Stein, C.1
Dep. Anesthesiology, Charité Campus Benjamin Franklin, Freie Universität
Berlin, Germany
2
Mouse Biology Unit, EMBL Monterotondo, Italy
1

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Opioid treatment of pain is limited by adverse side effects in thecentral nervous system (CNS). A better understanding of themechanisms underlying opioid analgesia mediated outside of the
CNS would help to develop opioids acting at peripheral sitesof tissue injury without centrally mediated unwanted effects. In
the CNS, activation of opioid receptors inhibits voltage-gatedcalcium (Ca2+)-channels and activates G protein-coupled inwardly
rectifying potassium (GIRK) channels through binding of G-protein
GEJVXEXQLWV%RWKHYHQWVUHGXFHPHPEUDQHH[FLWDELOLW\+RZHYHUVWXGLHVRQ*,5.FKDQQHOH[SUHVVLRQDQGIXQFWLRQLQSHULSKHUDOQHXURQVDUHVFDUFHDQGKDYHSURGXFHGFRQÁLFWLQJUHVXOWV+HUH

www.dcconferences.com.au/inrc2013/

we report that GIRK channels in sensory dorsal root ganglion(DRG) neurons are crucially involved in the generation of opioid
antinociception.
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*,5.VXEXQLWVLQ'5*RIQDwYHPLFH+RZHYHUH[SUHVVLRQRI
mRNA and protein of GIRK-1 and -2 was prominent in DRG ofnaïve rats. We detected GIRK currents after mu-opioid receptor
activation by DAMGO in rat but not in mouse DRG neurons. We
JHQHUDWHGWUDQVJHQLFPLFHH[SUHVVLQJD)ODJ*,5.FRQVWUXFW
selectively in DRG neurons by use of a Nav1.8 promoter.
DAMGO evoked GIRK currents in DRG neurons isolated fromthese Nav1.8-GIRK2 mice but not from wildtype littermates. We
DVVHVVHGKRZH[SUHVVLRQRI*,5.DIIHFWVQRFLFHSWLYHEHKDYLRU
by measuring hindpaw withdrawal in animals with painful hindpaw
LQÁDPPDWLRQ/RFDOLQMHFWLRQRI'$0*2DEROLVKHGWKHWKHUPDO
and mechanical hypersensitivity in Nav1.8-GIRK2 mice but not inwildtype littermates.
Our data show that GIRK channels in DRG neurons are crucial
for generation of peripheral opioid antinociception and that these
PROHFXODUPHFKDQLVPVPD\H[SODLQVSHFLHVGLIIHUHQFHV7KHVH
ÀQGLQJVVKRXOGEHFRQVLGHUHGLQGUXJGHYHORSPHQWDQGLQEDVLFDQG
clinical pain research.

72///,.(5(&(3725&+$1*(623,2,'
LIGAND BINDING IN BRAIN HOMOGENATES*Thomas, J.1, Somogyi, A.A.2, Hutchinson, M.R.3
Discipline of Pharmacology, University of Adelaide, Australia, 3Discipline of Physiology, University of Adelaide, Australia

1, 2

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In the infancy of opioid research, attention was focused directly
toward the stereoselective receptors that were discovered tobe critical for opioid analgesic responses. However opioids can
SDUDGR[LFDOO\LQFUHDVHSDLQVHQVLWLYLW\LQKXPDQVDQGURGHQWV
leading to hyperalgesia and tolerance. Work by the Hutchinson
and Watkins group has demonstrated that genetic removal of
LPPXQHUHFHSWRUVVXFKDVWROOOLNHUHFHSWRU 7/5 VLJQLÀFDQWO\reduces the development of hyperalgesia and tolerance
whilst simultaneously increasing anti-nociception. It is thoughtthat opioids bind non-stereoselectively to TLR4 leading to a
QHXURLQÁDPPDWRU\UHVSRQVHZLWKLQWKH&16FRPSURPLVLQJ
opioid-induced analgesia and contributing to various unwanted
DFWLRQV7KHH[LVWLQJGDWDRQWKHNLQHWLFVRIQRQVWHUHRVHOHFWLYH
opioid binding is limited and requires further investigation.
Wildtype (WT) and TLR4KO mouse brains were harvested
as required and homogenised using the gentleMACS™ Octo
Dissociator. Homogenates were diluted to 10mg/mL and [3H](-)
QDOR[RQHDGGHGWRQ0P/DOLTXRWVZHUHYDFXXPÀOWHUHG
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1DOR[RQHGLVVRFLDWLRQZDVLQLWLDWHGE\DGGLQJ0RIFROGGUXJ
Data was analysed using non-linear regressions. Our pilot data has
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NLQHWLFVLQ7/5.2 [VHFQ  PRXVHEUDLQKRPRJHQDWHVFRPSDUHGZLWK:7PRXVHEUDLQKRPRJHQDWHV [VHF
Q  7KLVZDVUHSUHVHQWHGE\IDVWHU  QDOR[RQHDVVRFLDWLRQLQ
the genetic absence of TLR4 compared to wildtype conditions
3 +RZHYHUXQH[SHFWHGO\ZHGLGQRWREVHUYHDFKDQJH
3! LQWRWDO>+@  QDOR[RQHELQGLQJEHWZHHQWKH:7 
DPM +/- 847 n=3) and TLR4KO (13895 DPM +/- 1169 n=3) brain
homogenates. These results further suggest an involvement of TLR4
LQRSLRLGSKDUPDFRORJ\EXWXQGHUOLHWKHFRPSOH[LW\RIWKLV7/5
opioid interaction.

43

POSTER ABSTRACTS

%,$6('6,*1$//,1*2)(1'2*(1286
23,2,'6$77+(085(&(3725*Thompson, G., Christopoulos, A. and Canals, M.
Monash Institute of Pharmacological Sciences, Drug Discovery Biology
Laboratory, Parkville,Victoria, Australia

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The endogenous opioid system, with multiple ligands targeting
WKHVDPHUHFHSWRUPD\UHSUHVHQWDQDWXUDOH[DPSOHRIIXQFWLRQDO
selectivity or signalling bias, where different ligands binding to
the same receptor generate different receptor conformations
linked to distinct signaling pathways and a particular physiologicalresponse. Understanding these mechanisms can provide essential
information on how opioid receptors regulate nociception andother physiological functions, including gastrointestinal motility, andmay lead to development of pain therapies that preserve potent
analgesia while minimising side effects, such as constipation. This
study aims to elucidate cellular and molecular mechanisms elicitedby endogenous opioids both in the central and enteric nervous
V\VWHPVDQGWRLQYHVWLJDWHWKHH[LVWHQFHRIIXQFWLRQDOVHOHFWLYLW\LQthe endogenous opioid system.
4XDQWLÀFDWLRQRIVLJQDOOLQJELDVRIDUDQJHRIHQGRJHQRXVRSLRLGV
at the Mu opioid receptor (MOR) has been performed in a
VLPSOHFHOOXODUPRGHOWRREWDLQXQLTXHOLJDQGDFWLYLW\SURÀOHVRU´ÀQJHUSULQWVµIRUWKHVHOLJDQGV7KHDELOLW\RIHDFKOLJDQGWRDFWLYDWH
multiple signal transduction pathways was measured; including G
protein activation, inhibition of adenylyl cyclase (AC), activation of
H[WUDFHOOXODUUHJXODWHGNLQDVHV (5.V UHFUXLWPHQWRIE-Arrestinsand receptor internalisation. The bias between each of these
SDWKZD\VZDVTXDQWLÀHGE\WKHDSSOLFDWLRQRIDQRYHODQDO\WLFDO
method based on the operational model of agonism. We found a
QXPEHURIHQGRJHQRXVRSLRLGVWKDWH[KLELWVLJQDOOLQJELDVDWWKH
MOR. As signalling through GPCRs is highly dependent upon the
FHOOXODUV\VWHPWKHOLJDQGDFWLYLW\SURÀOHVRIWKHVHHQGRJHQRXV
opioids were then validated in more physiological relevant cellular
V\VWHPV6SHFLÀFDOO\TXDQWLÀFDWLRQRIVLJQDOOLQJELDVKDVEHHQ
performed in a neuronal cell line, and currently we are generating
OLJDQGDGGLWLRQDODFWLYLW\SURÀOHVIRUELDVHGHQGRJHQRXVRSLRLGVLQ
primary dorsal root ganglia (DRG) and enteric neurons. This work
ZLOOHVWDEOLVKZKHWKHUVLJQDOOLQJELDVE\HQGRJHQRXVRSLDWHVH[LVWV
in primary DRG and enteric neurons.

'(9(/230(172)0,;('()),&$&<025'25/,*$1'6$1'02',),&$7,21
72,03529(%/22'%5$,1%$55,(5
3(1(75$7,21$QDQG-31<HRPDQV/13RUWHU%DUUXV951, Jutkiewicz,
E.M.2, *Traynor, J.R.2, Mosberg, H.I.
1
Department of Medicinal Chemistry, and (2) Department of Pharmacology,
University of Michigan, Ann Arbor, MI, 48109, USA.

0RQGD\²SP
It has been demonstrated that the co-administration of a mu
opioid receptor (MOR) agonist with a delta opioid receptor'25 DQWDJRQLVWSURGXFHVWKHH[SHFWHG025PHGLDWHG
analgesia, but displays reduced negative side effects, in particulara reduced level of tolerance and dependence, features that limit
the clinical use of opioid analgesics. However, combining two
GUXJVZLWKGLIIHUHQWVHOHFWLYLWLHVDQGHIÀFDFLHVKDVFRQVLGHUDEOH
disadvantages related to potential diverse pharmacokinetic
properties of the two chemical entities. Therefore we, and others,
have taken a multifunctional approach by combining both MOR
agonism and DOR antagonism in a single molecule. We have
VXFFHVVIXOO\JHQHUDWHGDVHULHVRIPL[HGHIÀFDF\F\FOLFSHSWLGHVusing our previously published homology models of the opioid
receptors to guide ligand design. These peptides when tested invitroGLVSOD\HGORZQDQRPRODUDIÀQLW\IRU025DQG'25ZLWK
moderate selectivity over the kappa opioid receptor (KOR). This
VHULHVRIOLJDQGVDOVRGLVSOD\HG025HIÀFDF\VLPLODUWRPRUSKLQH
and reduced DOR activation, with some compounds showing a
FRPSOHWHODFNRIHIÀFDF\DW'25$VORZELRDYDLODELOLW\LVW\SLFDO
of peptides, we installed a C-terminal E-glucoserine, in which
WKHVLGHFKDLQK\GUR[\ORIVHULQHLVFRYDOHQWO\2OLQNHGWRD
EJOXFRVHRQWRWKHPL[HGHIÀFDF\SHSWLGHVFDIIROGZLWKWKHPRVW
appropriate in vitro characteristics. This is a means of improving
blood-brain barrier penetration and increasing the metabolicstability. These E-glucoserine derivatives were assessed for in vivo
activity and shown to produce dose- dependent antinociception
after peripheral administration. This demonstrates that although
peptides are not usually thought of as “druggable” molecules,
WKHVHOLJDQGVDUHYLDEOHWRROVIRUH[SORULQJWKHUDSHXWLFSHSWLGH
analgesics as well as potentially developing molecules with reduced
dependence liability. Supported by RO1 DA-03910.

)XWXUHZRUNZLOOH[DPLQHWKHSK\VLRORJLFDORXWFRPHRIELDVHG
signalling by endogenous opioids by using animal models for
SDLQDQGJDVWURLQWHVWLQDOPRWLOLW\7KHVHH[SHULPHQWVZLOOHQDEOH
OLQNVEHWZHHQGLVWLQFWVLJQDOOLQJSURÀOHVDQGVSHFLÀFSK\VLRORJLFDO
responses in the intestine and spine to be established, providing
new information essential for the design opioids with effective
analgesia and less gastrointestinal side effects.

44International Narcotics Research Conference 2013

POSTER ABSTRACTS

22. A NOVEL MECHANISM FOR RESTRAINT
675(66,1'8&('&2&$,1(5(/$36(25(;,1,1'8&('(1'2&$11$%,12,'5(752*5$'(DISINHIBITION IN THE VENTRAL TEGMENTAL
AREA* Tung, L.W.1, Lu, G.L.1&KDQJ/<1/HH<64<X/6, Lee,
H.J.3,Teng, S.F.3, Hwang, L.L.4,5, Hung, M.S.7, Mackie, K.8,
=LPPHU$9 and Chiou, L.C.1,2,3
Grad Inst Pharmacol, 2 Grad Inst Brain and Mind Sciences, 3 Dept
3KDUPDFRO&RO0HG1DWLRQDO7DLZDQ8QLYHUVLW\7DLSHL7DLZDQ4 Grad Inst
Biomed Sci, 5'HSW3K\VLRO7DLSHL0HGLFDO8QLYHUVLW\7DLSHL7DLZDQ6 Inst%HKDY0HG&RO0HG1DWLRQDO&KHQJ.XQJ8QLYHUVLW\7DLQDQ7DLZDQ7
Inst Biotech Pharmaceut Res, National Health Research Institutes, Zhunan,
0LDROL7DLZDQ8 Gill Center and Dept Psychol Brain Sci, Indiana University,%ORRPLQJWRQ,QGLDQD86$9 Inst Mol Psychiat, University of Bonn, Bonn,
Germany.
1

7XHVGD\²SP$FWLYDWLRQRIK\SRWKDODPLFRUH[LQQHXURQVWKDWSURMHFWWRWKH
ventral tegmental area (VTA) is associated with the reinstatementof reward seeking, while its mechanism(s) remain unclear. Here,
we reveal a novel mechanism for stress-induced cocaine seeking,
LQYROYLQJRUH[LQLQGXFHGHQGRFDQQDELQRLG H&% VLJQDOLQJLQ97$
GRSDPLQHQHXURQV2UH[LQ$LQKLELWHG*$%$HUJLFWUDQVPLVVLRQ
in dopamine neurons of VTA slices via a presynaptic mechanism.
This effect was antagonized by OX1 (SB 334867) and CB1 (AM251), but not OX2 (Compound 29), antagonists, mimicked by a
CB1 agonist (WIN 55,212-2) and prevented by internally applied
GDP-ES (a G-protein inhibitor) and a phospholipase C inhibitor(U73122). JZL 184, an inhibitor of monoacylglyceral lipase thathydrolyses 2-arachidonoylglycerol (2-AG), an eCB, potentiated
DQGSURORQJHGWKHHIIHFWRIRUH[LQ$LQ97$VOLFHV&RQVLVWHQWO\WKHHIIHFWRIRUH[LQ$ZDVSUHYHQWHGE\WHWUDK\GUROLSVWDWLQDQ
inhibitor of diacylglycerol lipase, a 2-AG synthesizing enzyme. In
a conditioned place preference (CPP) test in mice, a 30 minrestraint stress, which increased the number of c-Fos-containing
RUH[LQQHXURQVLQWKHK\SRWKDODPXVDQGRUH[LQ$OHYHOVLQWKH97$
VLJQLÀFDQWO\UHLQVWDWHGH[WLQJXLVKHGFRFDLQH&337KLVDFXWHVWUHVV
induced cocaine reinstatement was prevented by SB 334867 or
AM 251, and was abolished in CB1 receptor-knockout mice. It wasalso prevented by a centrally applied CRF antagonist. These results
VXJJHVWWKDWDFXWHUHVWUDLQWVWUHVVDFWLYDWHVK\SRWKDODPLFRUH[LQ
QHXURQVWRUHOHDVHRUH[LQVZKLFKWKHQDFWLYDWHSRVWV\QDSWLF2;
receptors on VTA dopaminergic neurons, through a Gq-protein
coupled phospholipase C and diacylglycerol lipase enzymaticcascade, generating 2-AG. This eCB then retrogradely inhibits
GABA release through presynaptic CB1 receptors, leading todisinhibition of VTA dopamine neurons and cocaine seeking
behavior.

63,1$/$&7,2162)75,37$16$1'
23,2,'6,1$1,1)/$00$725<3$,167$7(
Jeong, H.J., Mitchell,V. and *Vaughan, C.
Pain Management Research Institute, Kolling Institute, Northern Clinical
School, University of Sydney at Royal North Shore Hospital, St Leonards,
NSW 2065, Australia.

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Triptans, such as sumatriptan, are 5-HT1B/D agonists which
are usually associated with the treatment of migraine. There is,however, evidence that triptans act within the spinal cord to
UHGXFHQRQFUDQLDOLQÁDPPDWLRQLQGXFHGVRPDWLFSDLQ 1LNDLHW
al. 2008 Pain 139:533-40). In the present study, we compared thespinal actions of opioids and triptans in a rat model of hindpaw
LQÁDPPDWLRQDQGH[DPLQHGWKHLUFHOOXODUDFWLRQVZLWKLQWKH

www.dcconferences.com.au/inrc2013/

OXPEDUVSLQDOFRUG$OOH[SHULPHQWVZHUHDSSURYHGE\WKH5R\DO
North Shore Hospital ACEC, and were performed on untreatedSprague-Dawley rats (naïve animals), or on rats 3 days after
intraplantar injection of complete Freund’s adjuvant (CFA animals).
Intrathecal injection of sumatriptan, the 5-HT1A agonist R(+)-8OH-DPAT and DAMGO all reduced hindpaw mechanical allodynia
in CFA animals. By contrast, DAMGO and R(+)-8-OH-DPAT, but
QRWVXPDWULSWDQUHGXFHGDFXWHWKHUPDO DQGWRDOHVVHUH[WHQWmechanical) pain in naïve animals. In lumbar spinal slices from naïve
and CFA animals, sumatriptan, R(+)-8-OH-DPAT and DAMGO allUHGXFHGWKHDPSOLWXGHRIDIIHUHQWHYRNHGH[FLWDWRU\SRVWV\QDSWLF
currents (EPSCs) in lamina I/II neurons. The inhibition of evoked(36&VSURGXFHGE\VXPDWULSWDQ YHUVXV DQG
'$0*2 YHUVXV ZDVKRZHYHUJUHDWHULQVOLFHV
from CFA compared to naïve animals. Furthermore, the 5-HT1B
agonist CP93129 inhibited evoked EPSCs in slices from CFA, but
QRWQDwYHDQLPDOV YHUVXVLQKLELWLRQ 7KH+7'
agonist PNU109291 had no effect on evoked EPSCs in lumbar
VOLFHVHLWKHUDQLPDOJURXS7KHVHÀQGLQJVVXJJHVWWKDWLQÁDPPDWLRQ
induces a functional upregulation of opioid and 5-HT1B receptor
systems in spinal lumbar pain pathways. Supported by Australian
NH&MRC (632546).

'(6,*16<17+(6,6$1'3+$50$&2/2*<2)23,2,''(/7$$*21,676:,7+$129(/)81'$0(17$/2;$=$75,&<&/2'(&$1(
6758&785(:DWDQDEH<1, 2, Hayashida, K.2, Saito, D.1,Takahashi,T.1,
Sakai, J.1, Nakata, E.1, Kanda,T.1, Iwai,T.2, Hirayama, S.2,
Nemoto,T.2, Fujii, H.2<DPDNDZD71 and Nagase, H.2
Discovery Research Laboratories, Nippon Chemiphar Co. Ltd., Saitama,
Japan and 2 School of Pharmacy, Kitasato University,Tokyo, Japan
1

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The GRSLRLGUHFHSWRULVH[SHFWHGWREHDQDWWUDFWLYHGUXJ
target without morphine-like side effects, but there is still no
G receptor selective agonist launched as a therapeutic agent.
For the past 20 years, although quite a lot of research for thesynthesis and pharmacology of G receptor agonists have beenreported, most of them are SNC-80 analogues which possess adiarylmethylpiperazine or a similar structure.
We have recently reported the synthesis of the morphinan
GHULYDWLYHVZLWKDQRYHOR[D]DWULF\FORGHFDQHVNHOHWRQ[1], [2], which
is a new structural class of G receptor agonists. Representative
FRPSRXQG6<.VKRZHGKLJKDIÀQLW\DQGPRGHUDWHVHOHFWLYLW\
for the GUHFHSWRU+RZHYHUWKLVFRPSRXQGH[KLELWHGORZDJRQLVWLF
activity in cAMP functional assay and only a weak analgesic effect inmice acetic acid writhing test.
In the present study, we postulated that the low agonistic activities
RIWKHR[D]DWULF\FORGHFDQHGHULYDWLYHVZRXOGPDLQO\UHVXOWIURPWKHH[LVWHQFHRIDSRVVLEOHDFFHVVRU\VLWHZKLFKLVDQH[WUD
structural part and interferes with a suitable conformationalchange of the G receptor bound to a GDJRQLVW LQGXFHGÀW 
Based on this hypothesis, we designed and synthesized derivatives
ZLWKRXWWKHSRVVLEOHDFFHVVRU\VLWHGLR[DPWK\OHQHPRLHW\$V
ZHH[SHFWHGWKHVHGHULYDWLYHVVKRZHGKLJKDIÀQLWLHVDQGIXOO
agonistic activities for the G receptor. Further optimization of thesestructures gave the improvement of analgesic activities. In our
ongoing studies, elucidation of the structure-activity relationships
RIWKHVHR[D]DWULF\FORGHFDQHGHULYDWLYHVZLOOSURYLGHQHZ
therapeutic agents.
[1] Hayashida, K.; Fujii, H.; Hirayama, S.; Nemoto, T.; Nagase, H. Tetrahedron,
2011, 67, 6682-6688.
[2] 8th AFMC International Medicinal Chemistry Symposium, Dec. 2011,
Japan

45

POSTER ABSTRACTS

25.THE ROLE OF DIFFERENT
3+263+25</$7,216,7(6,15$3,''(6(16,7,=$7,212)7+(23,2,'
5(&(3725<RXVXI$1, Miess, E.2, Schulz, S.2, Christie, M.J.1
Discipline of Pharmacology, University of Sydney, NSW, Australia,
2
Institute of Pharmacology and Toxicology, Jena University Hospital,
Friedrich Schiller University Jena, Jena Germany
1

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Tolerance and addiction to opioids are serious clinical andsocial problems that result in part from the loss of µ-opioid
receptor (MOR) function when activated by different opioids.
3KRVSKRU\ODWLRQRIVSHFLÀFUHVLGXHVLQWKH&WHUPLQDOGRPDLQRI
MOR is thought to be a key step in desensitization, endocytosis
and loss of MOR function. Sequential phosphorylation of S375
DORQJZLWKWKHÁDQNLQJUHVLGXHV77DQG7DUHUHTXLUHG
for endocytosis (Just et al., Mol Pharmacol (2013) 83:633-639)
but their role in rapid desensitization of MOR is unknown. The
DLPRIWKLVVWXG\ZDVWRGHWHUPLQHWKHLQÁXHQFHRI&WHUPLQDO
phosphorylation sites on rapid desensitization of MOR. Wild typemMOR, 3S/T-A (S363A, S370A, S375A), 6 S/T-A (S363A, S370A,
S375A, T376A, T379A, T383A) and 11S/T-A (T354A, S355A, S356A,
T357A, S363A, S370A, S375A, T376A, T379A, T383A, T394A)
ZHUHH[SUHVVHGVWDEO\LQ$W7FHOOV8VLQJSHUIRUDWHGSDWFKFODPSUHFRUGLQJZHH[DPLQHGWKHHIIHFWVRI025DFWLYDWLRQdesensitization and re-sensitization using activation of GIRKFKDQQHOVE\DVXEPD[LPDOFRQFHQWUDWLRQRIPHWHQNHSKDOLQ nM) to measure receptor activity and somatostatin (100 nM)
coupling to native SSTR2 (or sst2 according to IUPHAR) to
determine heterologous desensitization. MOR desensitization and
UHVHQVLWL]DWLRQSURGXFHGE\PLQH[SRVXUHWRPHWHQNHSKDOLQ
(10 µM) at 37oC did not differ from wild type for 3S/T-A or
6S/T-A but desensitization was abolished in the 11S/T-A mutant.
Desensitization, when detected, was largely homologous. Because
3S/T-A suppresses, and 6S/T-A abolishes MOR endocytosis, theseÀQGLQJVVXJJHVWWKDWKRPRORJRXVGHVHQVLWL]DWLRQFDQRFFXU
independently of the phosphorylation and arrestin-dependent
mechanisms that drive endocytosis. However, C-terminal
phosphorylation sites are necessary for desensitization because
mutation of all C-terminal sites (11S/T-A) abolishes desensitization.

'(6(16,7,=$7,212)7+(P23,2,'
5(&(3725,1,17$&7$77&(//6,6&7(50,1$/3+263+25</$7,21(66(17,$/"
Santiago, M.1'X<32, Christie, M.2, Connor, M.1
The Australian School of Advance Medicine, Macquarie University1, 2109
and Department of Pharmacology, University of Sydney, 2006, NSW,
Australia

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Internalization of the µ-opioid receptor (MOR) is not essential
for receptor desensitization and resensitization, and inhibiting
internalization may even increase resensitization speed.
Phosphorylation of the MOR c-terminal is important forinternalization, however, the relationship between receptor
phosphorylation and desensitization is not as well established.
It has also been suggested that protein kinase C-mediatedphosphorylation of MOR is associated with desensitization
produced by morphine but not DAMGO. We investigated acutedesensitization of mouse MOR (MOR-WT) and mutant MOR
in which important phosphorylation sites were deleted in intact
AtT-20 cells. Heterologous desensitization was assessed using
somatostatin receptor responses.
MOR-WT and 3 c-terminus phosphorylation site mutants: AAApoint mutations of S363, T370, S375; TPD - all c-terminal Ser/Thr
46

to Ala and TSST – TSST motif and T394 backmutant of TPD werestably transfected in AtT20 cells. MOR activation of GIRK was
PHDVXUHGXVLQJ)/,35PHPEUDQHSRWHQWLDOG\HLQD)OH[6WDWLRQ7KHGHJUHHRIGHVHQVLWL]DWLRQZDVTXDQWLÀHGXVLQJVDWXUDWLQJ
agonist concentrations added after the desensitizing stimulus.
DAMGO and morphine hyperpolarized AtT20-MOR cells.
7UHDWPHQWZLWKWKH3.&DJRQLVW30$RUWKHXQVSHFLÀFSURWHLQ
kinase inhibitor staurosporine did not affect agonist potency ordesensitization. Morphine hyperpolarized AtT-20-MOR cells and
WKRVHH[SUHVVLQJWKH$$$73'DQG7667YDULDQWVZLWK(&V(nM) of 16±5, 31±5, 14±2 and 13±2 respectively. After 30 minutes
treatment with morphine (1µM), the response to a subsequentDGGLWLRQRI0PRUSKLQHZDVUHGXFHGE\ :7  $$$  73' DQG 7667 7KHUHVSRQVH
to somatostatin (1µM) 30 minutes after morphine (1µM) was
LQKLELWHGE\ :7  $$$  73' DQG
(TSST). Qualitatively similar data was obtained for DAMGO. The
time course of the development of homologous and heterologous
desensitization was similar.
Mutations of key phosphorylation sites in the C-terminal regionof mouse MOR did not dramatically affect agonist potency, andmorphine induced desensitization of MOR persists even after
complete deletion of phosphorylation sites on c-terminal. Thissuggests that other regions of the receptor may be involvedin acute agonist-induced inhibition of signalling. Interestingly
WKHUHLVDVLJQLÀFDQWGLIIHUHQFHLQWKHGHJUHHRIKHWHURORJRXV
desensitization of somatostatin responses following activation ofsome of the MOR phosphomutants, suggesting distinct mechanisms
at play.

27. CAMKIID CONTROLS THE BIOGENESIS OF/(70,&5251$6,123,2,'72/(5$1&(*+H<DQG:DQJ=
Department of Biopharmaceutical Sciences, Cancer Center, & Program for
Collaborative Research in the Pharmaceutical Sciences, University of Illinois,
Chicago, 60612, USA

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Emerging evidence suggests that microRNAs (miRNAs) – mediatedcellular adaptations are critical for drug addiction. We previously
reported that let-7 family miRNAs contribute to the developmentof opioid tolerance by targeting the µ opioid receptor. Chronic
PRUSKLQHWUHDWPHQWLQGXFHGDPDUNHGLQFUHDVHRIOHWH[SUHVVLRQwhich functionally correlated with the development of opioid
tolerance.
The aim of this study was to understand the mechanisms how
OHWLVUHJXODWHGE\RSLRLGV:HÀUVWGHWHUPLQHGWKHWUDQVFULSWLRQVWDWXVRIOHWDQGIRXQGWKDWWKHH[SUHVVLRQRISULPDU\OHWSULOHW UHPDLQHGXQFKDQJHGLQ6+6<<FHOOVWKDWZHUHWUHDWHG
with morphine (1 PM, for 48 h). In agreement with the in vitro
observation, morphine pellet implantation (one 75 mg morphinepellet/mouse, s.c.) did not alter the level of pri-let-7 in mouse
EUDLQIURQWFRUWH[UHJLRQ7KHVHÀQGLQJVVXJJHVWHGWKDWWKHUREXVWelevation of let-7 occurred at the post-transcriptional level. Of
interest, in the presence of KN93, inhibitor of Ca2+ /calmodulindependent protein kinase II (CaMKII), chronic morphine treatment
IDLOHGWRJHQHUDWHOHWXSUHJXODWLRQLQ6+6<<FHOOV:HIXUWKHU
determined whether inactivation of CaMKIID by T286A pointPXWDWLRQZRXOGDIIHFWOHWH[SUHVVLRQDQGRSLRLGWROHUDQFH
Indeed, antinociceptive tolerance was absent in CaMKIIDT286A
mutant mice. Meanwhile, the level of let-7 in CaMKIIDT286A mutant
mice was much lower than that in wild-type mice, and wasresistant to chronic morphine stimulation. Taken together, thesedata suggested that the activity of CaMKIID was required for thebiogenesis of let-7 in opioid tolerance.
International Narcotics Research Conference 2013

POSTER ABSTRACTS

',65837,212)7+($&7,1
&<726.(/(721'(/$<65(&29(5<2)08
23,2,'5(&(3725)520'(6(16,7,=$7,21,1
/2&86&2(58/(861(85216
Arttamangkul, S.1, Schulz, S.2 and Williams, J.T.1
1Vollum Institute, Oregon Health & Science University, Portland, Oregon, USA
2 Pharmacology, Jena University Hospital, Jena, Germany

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The actin cytoskeleton acts to support cellular structure, but it
also plays a role in GPCR signaling and regulation. The network
RIDFWLQÀODPHQWVEHQHDWKSODVPDPHPEUDQHKDVEHHQVKRZQWRFRQÀQHPXRSLRLGUHFHSWRUV 025 LQWRDFRPSDUWPHQWDQG
restrains global lateral diffusion properties. The present study
used latrunculin B (Lat B) to disrupt actin polymerization in
HEK293 cells or mouse locus coeruleus neurons (LC). The effectof Lat B on lateral diffusion of Flag-tagged MOR (Flag-MOR) in+(.FHOOVZDVVWXGLHGE\OLQHVFDQQLQJÁXRUHVFHQFHFRUUHODWLRQ
spectroscopy. In Lat B treated cells, the diffusion rate of Flag-MOR
was faster than control. The effect of Lat B on [Met5]enkephalin(ME)-induced receptor internalization was studied in brain slices of
ORFXVFRHUXOHXVQHXURQVIURPWUDQVJHQLFPLFHH[SUHVVLQJ)ODJMOR. In the Lat B treated cells the amount of internalized FlagMOR and the endosome size was increased compared to control
following treatment with ME (30 µM, 10 min). The desensitization
measured by the decline in the hyperpolarization induced by ME(30 µM) was not altered but there was only a partial recovery
from desensitization following the wash. The impaired recovery
from desensitization correlated with a reduction in the recycling
RIUHFHSWRUVEDFNWRWKHSODVPDPHPEUDQH([SHULPHQWVXVLQJ/&slices obtained from mutant S375A knock-in mice showed that thedesensitization and the recovery were not altered by Lat B. This
study suggests that the actin cytoskeleton plays a key role in the
UHFRYHU\IURPGHVHQVLWL]DWLRQE\UHJXODWLQJWKHUHFHSWRUWUDIÀFNLQJThe study is supported by NIH grant DA08163.

3+263+25</$7,212)0823,2,'
5(&(37255('8&('0253+,1($1$/*(6,$,1,1)/$00$725<3$,167$7($RNL<1, Mizoguchi, H.1, Watanabe, C.1<RQH]DZD$1,
Sakurada,T.2, Sakurada, S.1
1
Department of Physiology and Anatomy,Tohoku Pharmaceutical University,
Sendai, Miyagi, Japan 2 First Department of Pharmacology, Daiichi College of
Pharmaceutical Sciences, Fukuoka, Japan

0RQGD\²SP$QLQÁDPPDWRU\SDLQLVDFKURQLFSDLQLQGXFHGE\DOJHVLFRULQÁDPPDWRU\VXEVWDQFHV,WLVZHOOHVWDEOLVKHGWKDWPRUSKLQHLV
HIIHFWLYHDJDLQVWWKHWKHUPDOK\SHUDOJHVLDGXULQJLQÁDPPDWRU\pain state. However, we found that morphine is ineffective against
WKHPHFKDQLFDODOORG\QLDRQWKHLQÁDPPDWRU\SDLQVWDWH,QWKH
present study, the changes in the spinal mu-opioid receptors,
which is involved in the morphine analgesia, is investigated in
WKHLQÁDPPDWRU\SDLQVWDWH7RGHYHORSWKHLQÁDPPDWRU\SDLQcomplete Freund’s adjuvant (CFA) was injected i.pl to the hindSDZRIPDOHGG<PLFH7KHSURWHLQOHYHORIPXRSLRLGUHFHSWRUVZDVTXDQWLÀHGE\ZHVWHUQEORW0HFKDQLFDOSDLQWKUHVKROGDQG
DQDOJHVLFHIIHFWRIPRUSKLQHZHUHPHDVXUHGE\YRQ)UH\ÀODPHQW
test. The remarkable mechanical allodynia was observed after the&)$LQMHFWLRQRQLQÁDPHGVLGHEXWQRWQRQLQÁDPHGVLGH7KH

www.dcconferences.com.au/inrc2013/

analgesic effect of morphine injected s.c. was markedly decreased
bilaterally after the CFA injection. However, the protein levels
RIPXRSLRLGUHFHSWRUZHUHVLJQLÀFDQWO\GHFUHDVHGLQWKH'5*
RQO\RQLQÁDPHGVLGHDIWHU&)$LQMHFWLRQ7KHSUHVHQWUHVXOWVVXJJHVWWKDWWKHUHGXFWLRQRIPRUSKLQHDQDOJHVLDLQLQÁDPPDWRU\SDLQVWDWHRQLQÁDPHGVLGHUHÁHFWWKHGRZQUHJXODWLRQRIPX
opioid receptor, whereas another mechanism is also involved
LQWKHUHGXFWLRQRIPRUSKLQHDQDOJHVLDRQQRQLQÁDPHGVLGH
LQLQÁDPPDWRU\SDLQVWDWH,QWHUHVWLQJO\WKHUHGXFHGPRUSKLQH
analgesia after CFA injection was completely reversed on nonLQÁDPHGVLGHDQGSDUWLDOO\UHYHUVHGRQLQÁDPHGVLGHE\LQWUDWKHFDO
pretreatment with protein kinase C inhibitor, which was injected
DWWKHVDPHWLPHRIGHYHORSPHQWRILQÁDPPDWLRQE\&)$LQMHFWLRQ,QFRQFOXVLRQWKHUHGXFWLRQRIPRUSKLQHDQDOJHVLDRQLQÁDPHG
VLGHPD\PDLQO\UHÁHFWWKHGRZQUHJXODWLRQRIPXRSLRLGUHFHSWRUZKHUHDVWKHUHGXFWLRQRIPRUSKLQHDQDOJHVLDRQQRQLQÁDPHG
VLGHPD\PDLQO\UHÁHFWWKHLQDFWLYDWLRQRIPXRSLRLGUHFHSWRUE\phosphorylation.

7+('(/7$23,2,'5(&(3725
&2175,%87(6720253+,1(72/(5$1&(IN AN ANIMAL MODEL OF CHRONIC,1)/$00$725<3$,1
Beaudry, H. 1,2, Hipólito, L.2, Gupta, A.3, Devi, L.3, Gendron,
L.1, Morón Concepción, J.A.2
1
Dept. of Physiology and Biophysics, Université de Sherbrooke,
Sherbrooke, Canada, 2 Dept. of Anesthesiology, Columbia University
Medical Center, New York, USA, 3 Department of Pharmacology and
Systems Therapeutics, Mount Sinai School of Medicine, New York, USA.
7XHVGD\²SP
Opioids are well known for their robust analgesic effects. Although
they can activate 3 receptor subtypes (mu, delta, and kappa
opioid receptors; MOPRs, DOPRs, KOPRs), in vivo, commercial
opioids preferentially target MOPRs. Chronic activation of MOPr
is however accompanied by various unwanted effects such as
analgesic tolerance. Among other mechanisms, interactions
between MOPR and DOPR are thought to play an important rolein morphine-induced behavioral adaptations. Here, we investigated
the role and the regulation of DOPR and the MOPR/DOPR
heteromer formation during the development of morphineWROHUDQFHLQDQDQLPDOPRGHORIFKURQLFLQÁDPPDWRU\SDLQ:H
found that prolong morphine treatment, in the setting of chronic
SDLQLQGXFHGDQLQFUHDVHLQ'235H[SUHVVLRQDWWKHSRVWV\QDSWLF
density fraction in the dorsal horn of the spinal cord. In addition,
our results showed an increase in MOPR/DOPR heteromer
abundance in spinal cord dorsal horn and periaqueductal grayhomogenates. Finally, using behavioral approaches, we observedthat blockade of DOPR with the selective antagonist naltrindole(s.c. or i.t.) attenuates the development of morphine tolerance
in a dose-dependent manner. Altogether, our results suggest thattargeting the DOPR provides a valuable strategy to attenuate
the analgesic tolerance that develops after repeated morphine
DGPLQLVWUDWLRQLQWKHVHWWLQJRIFKURQLFLQÁDPPDWRU\SDLQFurther studies are needed to understand cellular and molecularmechanisms involved in this process.
This work was supported by Departmental start up funds to JAMC, by a CIHR
grant to LG and a FRQ-S fellowship to HB.

47

POSTER ABSTRACTS

(1'272;,1(1+$1&(0(172)
0253+,1(,1'8&('&21',7,21('3/$&(
35()(5(1&(*Sarkar, S.10DR;1,Vigorito, M.1,2, Chang, S.L.1,3
Institute of Neuroimmune Pharmacology, 2Department of Psychology, andDepartment of Biological Sciences, Seton Hall University, 400 South Orange
Avenue, South Orange, NJ 07079, USA
1
3

0RQGD\²SP
Because of its potent analgesic effects, morphine is widely usedin clinical pain management. However, it has a high potential
IRUDEXVHDQGDGGLFWLRQ,QWKLVVWXG\ZHH[DPLQHGWKHHIIHFWVRIWKHHQGRWR[LQOLSRSRO\VDFFKDULGH /36 RQWKHVHQVLWLYLW\WR
morphine in adult rats. Adult Fischer 344 rats were given two
intraperitonial (i.p.) injections of either LPS (250 µg/kg) or saline(vehicle), followed by an injection of either a higher dose of LPS(2 mg/kg) or saline after 24 h. After 72 h, the rats were given an
i.p. injection of either morphine (3.5 mg/kg) or saline every otherday during the 6 d of the conditioning phase of a conditioned
SODFHSUHIHUHQFH &33 H[SHULPHQW'XULQJWKHSRVWFRQGLWLRQLQJ
phase, the rats were assessed for their drug preference. In thegroups given LPS, there was a stronger preference for morphinecompared to the group that was given only saline, and there was asustained response to morphine as evidenced by greater than 20 d
RIH[WLQFWLRQ,QDSDUDOOHOVWXG\WDLOÁLFNODWHQFLHVZHUHPHDVXUHGWRHYDOXDWHWKHDQDOJHVLFHIIHFWVRIPRUSKLQHIROORZLQJH[SRVXUH
to LPS. The rats were pretreated with LPS (2 mg/kg) or saline,
followed by treatment with cumulative doses of morphine ranging
IURP²PJ.J7DLOÁLFNODWHQFLHVLQUHVSRQVHWRPRUSKLQH
ZHUHVLJQLÀFDQWO\ORQJHULQWKHUDWVJLYHQ/36FRPSDUHGWRWKRVH
given saline. Collectively, our data indicate that systemic infections
can increase the sensitivity to morphine both physiologicallyand behaviorally (partially supported by K02 DA016149 & RC2
AA019415 to SL Chang).

32. ADOLESCENT DRINKING AFFECTS(1'2*(128623,2,'6,1+,332&$0386
$1'$0<*'$/$,1'(3(1'(172)5($5,1*
CONDITION
3DOP6 Daoura, L., Roman, E. and Nylander, I.
Neuropharmacology, Addiction & Behaviour, Department of Pharmaceutical
Bioscience, Uppsala University, Uppsala, Sweden

7XHVGD\²SP
Early onset of alcohol drinking predicts increased risk for alcohol
use disorders (AUD) later in life but the mechanisms are notunderstood. A link between alcohol and endogenous opioids
are well described and it is known that endogenous opioids areaffected by early-life rearing conditions as evidenced by distinct
changes in the opioid networks in young and adult rats. However,
the effects of adolescent drinking on opioids are poorly described
and little is known whether these effects are dependent on
HDUO\OLIHH[SHULHQFHV7RRSWLPL]HDQGLQGLYLGXDOL]HSUHYHQWLRQ
and treatment strategies for AUD, a better understanding of thelink between the early-life environmental factors, ethanol drinking
and endogenous opioids is of importance. The aim was to study
interactions between rearing conditions and adolescent drinking
RQWKHRSLRLGSHSWLGHVG\QRUSKLQ% '<1% PHWHQNHSKDOLQ
Arg6Phe7 (MEAP) and beta-endorphin (BEND).
A maternal separation (MS) model was used to simulate different
rearing conditions. The pups were separated 360 min daily from
WKHGDPGXULQJWKHÀUVWWKUHHSRVWQDWDOZHHNV3XSVVXEMHFWHG
to 15 min MS were controls. After weaning, on postnatal day 34,
all rats were individually housed and given a free choice between
HWKDQRODQGZDWHURUZDWHURQO\IRUZHHNV(WKDQROZDV
available 24h on Mondays, Wednesdays and Fridays.
48

A main effect of ethanol was found with higher immunoreactive
LU OHYHOVRI'<1%LQWKHKLSSRFDPSXVDQGKLJKHULU0($3OHYHOVin the amygdala. A main effect of rearing condition was found in thepituitary gland and periaqueductal grey with lower ir BEND levelsin the MS360 rats. No interaction between rearing condition andethanol was observed. Previous studies have reported differences
in basal and ethanol-induced effects in adult rats subjected to MS15
and MS360. This study shows that adolescent drinking attenuatethese differences. It is possible that the individual housing had
a larger impact on opioid levels than the rearing environment
and therefore masked the previously seen differences in opioid
SHSWLGHV6LQFH'<1%LPSDLUVOHDUQLQJDQG0($3UHGXFHVDQ[LHW\like behaviour it is of particular interest to note that voluntary
DGROHVFHQWGULQNLQJWDUJHWVKLSSRFDPSDO'<1%DQG0($3LQWKH
amygdala.
Funded by SRA (SRA 2011-0056), the Swedish Medical Research Council (201261X-22090-01-3) and ERAB (EA 11-30).

6,1*/(+286,1*'85,1*$'2/(6&(1&(&$86($/7(5$7,216,1%$6$//(9(/62)
(1'2*(128623,2,'3(37,'(6Roman, E., *Granholm, L., Nylander, I.
Neuropharmacology, Addiction and Behaviour, Department of Pharmaceutical
Bioscience, Uppsala University, Sweden

0RQGD\²SP
7KHHQGRJHQRXVRSLRLGV\VWHP (26 H[HUWDYDULHW\RIHIIHFWVand one of its most noteworthy functions is the regulation of
QDWXUDOUHZDUGVOLNHIRRGVH[DQGVRFLDOLQWHUDFWLRQV'LVWXUEDQFH
of natural reinforcers can cause basal alterations in the EOS andconsequently contribute to an intrinsic vulnerability for different
neurobiological disorders, e.g. addiction and mood disorders.
Early in life, social interactions are important for the normal
development of the EOS. In this study, the aim was to investigate
the effects of social isolation (i.e. single housing with the possibility
to hear and smell other animals) during adolescence on basal levelsof endogenous opioids.
On postnatal day 22 (PND22) Sprague-Dawley pups were weaned
DQGUDQGRPO\DVVLJQHGLQWRWKUHHGLIIHUHQWH[SHULPHQWDOJURXSVlong social isolation (LSI), short social isolation (SS1) and controls.
$W31'WKHUDWVZHUHVDFULÀFHGE\GHFDSLWDWLRQ7KHDQLPDOV
in the LSI group were individually housed from PND22 until
PND29. The SSI rats were group housed until 30 minutes prior
to decapitation when they were singly housed in a separate cage.
The controls were group housed until decapitation. The brains
were dissected and trunk blood collected. Tissue levels of three
opioid peptides, Met-enkephalin-Arg6Phe7 (MEAP), dynorphin B'<1% DQGQRFLFHSWLQRUSKDQLQ)4 12)4 DVZHOODVVHUXP
corticosterone were analysed using radioimmunoassays.
7KH/6,UDWVKDGVWDWLVWLFDOO\VLJQLÀFDQWORZHULPPXQRUHDFWLYH LU 
0($3OHYHOVFRPSDUHGWRFRQWUROVLQK\SRWKDODPXVIURQWDOFRUWH[PHGLDOSUHIURQWDOFRUWH[VWULDWXPDP\JGDODVXEVWDQWLDQLJUDDQG
periaqueductal gray. Elevated levels of ir N/OFQ were seen inboth SSI and LSI compared to controls. None of the isolation
JURXSVGLIIHUHGLQLU'<1%OHYHOVFRPSDUHGWRFRQWURO7KHLU
corticosterone in SSI rats were increased compared to controls.
These results show that disturbance of social interactions during
DGROHVFHQFHLQGXFHFKDQJHVLQHQGRJHQRXVRSLRLGVDQGFRQÀUPthe importance of social behaviour for development during thissensitive time window. A deranged EOS may have long-term
consequences for later susceptibility to addiction and otherdisorders where the EOS is implied in pathogenesis.
Funded by SRA (SRA 2011-0056), the Swedish Medical Research Council (201261X-22090-01-3) and the ERAB (EA 11 30).

International Narcotics Research Conference 2013

POSTER ABSTRACTS5('8&('0,&5251$%&/867(5(;35(66,21,17+(18&/(86$&&80%(16,6,192/9(',17+(326775$16&5,37,21$/
02'8/$7,2181'(51(8523$7+,&3$,1*Hamada, A.1<DPDVKLWD$16XKDUD<1, Narita, M.1,
Tsuyama, J.2, Kuzumaki, N.2, Okano, H.2 and Narita, M.1
1
Dept. Pharmacol, Hoshi Univ. Sch. Med.Tokyo, Japan, 2 Dept. Physiol.,
Keio Univ. Sch. Med.Tokyo, Japan.
7XHVGD\²SP
1HXURSDWKLFSDLQLVWKHPRVWGLIÀFXOWW\SHRISDLQWRFRQWURODQG
patients lose their motivation for the purposive pursuit with adecrease in their quality of life. It has been widely recognized thatthe ascending anatomical dopamine projection from the ventral
tegmental area (VTA) to the nucleus accumbens (N.Acc.) is mostly
related to motivational functions and the reinforcing effects of
opioids. On the other hand, microRNAs (miRNAs) are small,noncoding RNA molecules that direct the post-transcriptional
VXSSUHVVLRQRIJHQHH[SUHVVLRQDQGSOD\DQLPSRUWDQWUROHLQregulating synaptic plasticity. MiRNAs have been shown to respondto various cellular stressors to play a role on psychiatric disorders,such as schizophrenia and autism. Thus, miRNAs are thought toplay a role in functions of the central nervous systems. In this study,
we found that sciatic nerve ligation induced a drastic decrease in
WKHH[SUHVVLRQRIPL5EDQGPL5LQ1$FFQHXURQV7KH
H[SUHVVLRQRI'1$PHWK\OWUDQVIHUDVHD '107D ZKLFKLV
WKHRQHRIWKHSUHGLFWHGWDUJHWVRIPL5EZDVVLJQLÀFDQWO\LQFUHDVHGLQWKH1$FFDWGD\VDIWHUVFLDWLFQHUYHOLJDWLRQ1H[W
ZHPLFURLQMHFWHGWKHPL5EH[SUHVVLRQOHQWLYLUXVLQWRWKH
N.Acc. to evaluate pain behaviors by sciatic nerve ligation. We
IRXQGWKDWLWVPLFURLQMHFWLRQSDUWLDOO\EXWVLJQLÀFDQWO\UHGXFHGthe thermal hyperalgesia and mechanical allodynia by sciatic nerve
ligation. The results of these analysis provide new insight into a
SRVWWUDQVFULSWLRQDOPRGLÀFDWLRQDWWKHEUDLQWKDWLVDFFRPSDQLHG
by a dramatic decrease in miR200b and miR429 along with thedysfunction of “mesolimbic motivation/valuation circuitly” undera neuropathic pain-like state. These phenomena may result in adecrease in DNMT3a in neurons of the N.Acc. under neuropathic
pain, which leads to the long-term transcription-silencing of several
genes.
35.THE TRANSLOCATION OF THE G23,2,'
5(&(372521&+2/,1(5*,&,17(51(85216,1
7+(675,$780*Heath, E.1, Chieng, B.1, Christie, M.2, Balleine. B.1
1
Brain and Mind Research Institute,The University of Sydney, NSW 2050
Australia, 2 Discipline of Pharmacology, University of Sydney, NSW, Australia

0RQGD\²SP
In many neuronal populations under baseline conditions the
G-opioid receptor (DOR) is constitutively retained in intracellularcompartments with only low levels of DOR present at the plasma
membrane. Under certain conditions intracellular DOR may beinserted into the cell membrane, increasing the levels of functional
DOR and enhancing the effects of DOR agonists. In the striatum,'25LVNQRZQWREHKLJKO\H[SUHVVHGRQFKROLQHUJLFLQWHUQHXURQV
(CIs), whose inhibitory function is believed to play a vital role in
the regulation of the striatum. In these neurons, DOR follows the
distribution pattern described above – with neurons containing
non-functional, intracellular pools of the receptor in addition to
H[SUHVVLQJVRPHUHFHSWRUDWWKHPHPEUDQH5HVHDUFKLQRXU
lab has demonstrated that, in the NACsh of DOR-eGFP mice,following Pavlovian conditioning DOR has translocated from thewww.dcconferences.com.au/inrc2013/

cytoplasm to the perisomatic membrane of CIs. The translocation
and activity of DOR in the NACsh is not required for Pavlovian
OHDUQLQJRUWKHH[SUHVVLRQRI3DYORYLDQEHKDYLRXUVEXWIRUWKH
H[SUHVVLRQRIRXWFRPHVSHFLÀF3DYORYLDQLQVWUXPHQWDOWUDQVIHU
(PIT). However, while this demonstrates a behavioural trigger for
DOR insertion, the molecular mechanisms of this translocation
event remain unknown. The present study investigates whether theneurotransmitter substance P (SP) may drive insertion of DOR
into the perisomatic membrane of CIs in the striatum. In the past,
SP has been controversially linked to DOR translocation in small
dorsal root ganglion neurons via a proposed interaction between'25DQG63OHDGLQJWRWKHLUFRWUDIÀFNLQJ7KLVPHFKDQLVPKDV
been challenged by later studies. Here, we propose an alternativeinteraction between the two molecules – namely that SP
signalling triggers the translocation of DOR. SP (300nm) was
administered to the striatum of DOR-eGFP mice both in vivo
and in vitro. Immunohistochemical staining for GFP and choline
acetyltransferase (ChAT) was performed on treated sections and'25H*)3UHFHSWRUDWWKHPHPEUDQHRI&,VZDVTXDQWLÀHG$SSOLFDWLRQRI63WRWKHGRUVDOPHGLDOVWULDWXP '06 VLJQLÀFDQWO\increased the amount of DOR-eGFP (p<0.01) at the perisomatic
membrane of CIs in the region. This demonstrates a novel
mechanism of DOR translocation which may contribute to the
behavioural effects of opioids in the striatum.

320&1(85216,17+(+<327+$/$086
02'8/$7(3(5,3+(5$/,0081()81&7,21*Ikegami, D.17DVDNL<1, Suzuki, M.2, Aoki, K.38H]RQR<2and Narita, M.1
1
Dept. Pharmacol., Hoshi Univ. Sch. Pharm. Pharmaceut. Sci., 142-8501,
Japan 2 Div. Cancer Pathophysiol., NCCRI, 104-0045, Japan 3 Div. Gene and
Immune Medicine., NCCRI, 104-0045, Japan

7XHVGD\²SP
7KHSDWKRSK\VLRORJLFDOVWDWHRIFDQFHUFDFKH[LDIUHTXHQWO\affects advanced cancer patients. The close association between
chronic illness such as cancer and the nutritional deterioration
does not only impair the quality of life but also increases the risk
for morbidity and mortality. On the other hand, E-endorphin,an endogenous opioid polypeptide primarily produced by the
hypothalamus, is known to have the ability to inhibit stresshormone production and produces analgesia, and enhances
innate immune function. E-Endorphin is a cleavage product of
pro-opiomelanocortin (POMC), which is also the precursors
hormone for ACTH and melanocyte stimulating hormone(D06+ :HLQYHVWLJDWHGSRVVLEOHFKDQJHVLQWKHH[SUHVVLRQRI
320&LQWKHK\SRWKDODPXVXQGHUWKHFDQFHUFDFKH[LDFDXVHGE\intraperitoneal inoculation of pancreatic cancer cells. At 3 weeks
DIWHUWXPRULQRFXODWLRQZKHQPLFHH[KLELWHGVOLJKWZHLJKWORVV
320&H[SUHVVLRQLQWKHK\SRWKDODPXVZDVVLJQLÀFDQWO\GHFUHDVHG
compared to that in control mice. Under these conditions,decrease in the macrophage colony-stimulating factor (M-CSF)
was found in the serum from the peritoneal metastasis model mice.
Furthermore, we found that the M-CSF was decreased in the
VHUXPIURPWKH320&NQRFNRXWPRXVH1H[WZHXVHG320&
FUHPRXVHZLWK$$9IRU320&QHXURQVSHFLÀFSKRWRVWLPXODWLRQ
The suppression of POMC neurons in the hypothalamus resulted
LQGHFUHDVHG0&6)LQWKHPRXVHVHUXP7KHVHÀQGLQJVVXJJHVW
that POMC neurons engage to activate peripheral immunity and
DQWLLQÁDPPDWRU\F\WRNLQHV

49

POSTER ABSTRACTS

,192/9(0(172)'(/7$23,2,'(5*,&
6<67(0,1$'8/71(852*(1(6,6*Iwasawa, C.1, Watanabe, M.1+DPDGD<1, Kuzumaki, N.1,2,
Narita, M.1, Sawamoto, K.3, Narita, M.1
1
Dept. Pharmacol., Hoshi Univ. Sch. Pham. Pharmaceut. Sci.,Tokyo, Japan 2
Dept. Physiol., Keio Univ. Sch. Med.,Tokyo, Japan 3 Dept. Dev. Regene. Biol.,
Nagoya City Univ., Aichi, Japan

0RQGD\²SP
Opioid analgesics and endogenous opioid peptides have a wide
range of physiologic and behavioral effects on pain perception,mood, motor control and autonomic function. The deltaopioidergic system has been recognized as a neurotransmitter
system that could be directly involved in emotionality andimmunity. Recently, we reported that a long-term blockade ofdelta-opioid receptor function using the selective delta-opioid
receptor antagonist naltrindole (NTI) induced astrogliosis. TheseÀQGLQJVVXSSRUWWKHFRQFHSWWKDWWKHGHOWDRSLRLGUHFHSWRUSOD\V
an important role in cell proliferation, gliogenesis and neurogenesis.
Neural stem cells capable of producing new neurons andastrocytes are maintained in a self-replicating state by cytokines
and neurotrophic factors. In the present study, we investigated
the role of the delta-opioidergic system in adult neurogenesis.
We found a dramatic increase of neural progenitor cells in the
hippocampal dentate gyrus of mice that were chronically treatedwith the selective delta-opioid receptor agonist. In addition,
we showed that enriched environment-induced hippocampal
QHXURJHQHVLVZDVLQKLELWHGE\WUHDWPHQWZLWK17,7KHVHÀQGLQJV
suggest that the endogenous delta-opioidergic system could
facilitate hippocampal neurogenesis. We are currently investigating
the possible involvement of delta-opioid systems in the migrationof immature neuronal cells in the brain.

',))(5(17,$/())(&762)0253+,1($1'
0(7+$03+(7$0,1(217+($&7,9$7,21
OF MESOLIMBIC AND NIGROSTRIATAL'23$0,1(5*,&6<67(06,ZDVH<Mori,T., Saeki,T., Iwata, N., Murata, A., Shibasaki,
M., Suzuki,T.
Dept Toxicology, Hoshi University School of Pharmacy and Pharmaceuticalsciences,Tokyo 142-8501, Japan

7XHVGD\²SP
Methamphetamine and morphine induce several behavioral
changes mediated through the activation of dopaminergic system;high doses of methamphetamine induce a decrease of locomotor
activity accompanied by stereotyped behavior in rats and mice,
whereas high doses of morphine induce hyperlocomotion inmice and hypolocomotion in rats. We hypothesized here that
distinct phenotype of behavior induced by methamphetamineand morphine would result from differential activation for themesolimbic and nigrostriatal dopaminergic systems. Therefore,
ZHÀUVWO\PHDVXUHGGRSDPLQHUHOHDVHIURPQXFOHXVDFFXPEHQV
and striatum simultaneously induced by methamphetamine
or morphine using in vivo microdialysis in rats. In the present
study, methamphetamine potently increased the release of
dopamine from both nucleus accumbens and striatum, whereasmorphine increased the release of dopamine from only nucleus
accumbens. It is known that inhibition of GABA release into theventral tegmental area, which is the cell body of the mesolimbic

50

dopaminergic system, is critically involved in the release of
GRSDPLQHIURPWKHQXFOHXVDFFXPEHQVE\PRUSKLQH:HQH[W
H[DPLQHGWKHUHOHDVHRI*$%$LQWRWKHYHQWUDOWHJPHQWDODUHD
and substantial nigra compacta, the cell body of nigrostriatal
dopaminergic system by morphine using microdialysis. As we
H[SHFWHGLQKLELWLRQRI*$%$UHOHDVHLQGXFHGE\PRUSKLQH
was only observed in the ventral tegmental area, but not in thesubstantial nigra compacta. Furthermore, the ratio between
PRSLRLGUHFHSWRUVYV*$'P51$ZDVVLJQLÀFDQWO\KLJKHU
in the ventral tegmental area than those in the substantial nigra
FRPSDFWD7KHVHGLIIHUHQFHVPD\LQÁXHQFHWKHGLIIHUHQWLDOUHOHDVLQJ
pattern of dopamine from the nucleus accumbens and striatum
by morphine. Furthermore, differential effects of morphine andmethamphetamine on the activation of mesolimbic and nigrostriatal
dopaminergic systems result in the induction of differential
phenotype of behavior in rodents.

,192/9(0(172)6,*0$5(&(2725
&+$3(521(217+((;35(66,212)
:,7+'5$:$/6,*16,10253+,1('(3(1'(170,&(*Ohya, J., Mori,T., 8]DZD16XJL\DPD.6DLWRK<Shibasaki, M., Suzuki,T.
Dept Toxicology, Hoshi University School of Pharmacy and Pharmaceuticalsciences,Tokyo 142-8501, Japan

0RQGD\²SP
It is known that withdrawal syndromes after the cessation of
P-opioid receptor agonists remains an obstacle in the clinicaltreatment of pain, and free radical formation is taking a part in
WKHH[SUHVVLRQRIZLWKGUDZDOVLJQVLQPRUSKLQHGHSHQGHQWPLFH
5HFHQWÀQGLQJVVXJJHVWWKDWVLJPDUHFHSWRUFKDSHURQHKDVD
prominent role to maintain the homeostasis against several stress
LQFOXGLQJR[LGDWLYHVWUHVVE\TXHQFKLQJHIIHFWIRUIUHHUDGLFDO
Therefore, the present study was designed to investigate theinvolvement of sigma-1 receptor chaperone as well as free radical
RQWKHH[SUHVVLRQRIZLWKGUDZDOVLJQVLQPRUSKLQHGHSHQGHQW
mice. Morphine was injected by slow release emulsion, andZLWKGUDZDOVLJQVZHUHSUHFLSLWDWHGE\WKHLQMHFWLRQRIQDOR[RQH
day after the slow release emulsion administration of morphine.
:LWKGUDZDOVLJQVSUHFLSLWDWHGE\QDOR[RQHLQPRUSKLQHGHSHQGHQW
mice, especially body-weight loss and diarrhea, were almost
completely suppressed by the sigma-1 receptor antagonist NE100
and the free radical scavenger fullerene. Furthermore, we also
found that protein level of sigma-1 receptor chaperone werepotently upregulated in the ascending colon. These results suggestthat sigma-1 receptor chaperone as well as formation of free
UDGLFDOSOD\VDVLJQLÀFDQWUROHLQWKHH[SUHVVLRQRIZLWKGUDZDOVLJQVLQPRUSKLQHGHSHQGHQWPLFH3UHYLRXVÀQGLQJVKRZHGWKDW
5-HT plays a role in the diarrhea during the withdrawal state
LQPRUSKLQHGHSHQGHQWUDWV7KHUHIRUHZHQH[WH[DPLQHGWKH
interaction between 5-HT and free radical or sigma-1 receptor
chaperone. In the present study, 5-HT-induced diarrhea was
suppressed by fullerene, however NE100 did not affect 5-HTLQGXFHGGLDUUKHD7KHVHÀQGLQJVLQGLFDWHWKDWIUHHUDGLFDOVFDYHQJHU
and sigma-1 receptor antagonist could independently suppress
withdrawal diarrhea in morphine-dependent rodents.

International Narcotics Research Conference 2013

POSTER ABSTRACTS23,2,'(5*,&0(&+$1,60,6/2&$/,=('IN THE DOWNSTREAM OF D7 NICITINIC$&(7</&+2/,1(5(&(3725%87127D4E2, IN
7+(&(175$/1(592866<67(0*Kishioka, S., Kiguchi, N., .RED\DVKL<Saika, F., Wakida, N.,<DPDPRWR&
Department of Pharmacology,Wakayama Medical University School of
Medicine,Wakayama 641-0012 Japan

7XHVGD\²SP
Nicotine is a neuronal stimulant and produces various
pharmacological effects through the activation of nicotinic
acetylcholine receptors (nAChRs) in the central nervous system;
IRUH[DPSOHDQWLQRFLFHSWLRQDQGVHUXPFRUWLFRVWHURQH 6&6 
increase. Nicotine also has a physical dependence liability. And
there is a growing evidence suggesting that nicotine effects, at leastin part, elicit through the activation of opioidergic system. On theother hand, it is well-known that two nAChR subtypes, i.e., D7 andD4EQ$&K5VDUHH[SUHVVHGLQWKHFHQWUDOQHUYRXVV\VWHP7KH
DLPRIWKLVH[SHULPHQWLVWRFODULI\WKHQ$&K5VXEW\SHZKLFKZDV
coupled with opioidergic system. Nicotine-induced antinociception
ZDVHYDOXDWHGE\WDLOSLQFKWHVWDQG6&6OHYHOZDVTXDQWLÀHGE\ÁXRURPHWULFDODVVD\XVLQJ,&5PDOHPLFH)RUWKHHVWLPDWLRQRI
nicotine withdrawal, we evaluated SCS increase as an indicator,
because we have reported that the magnitude of SCS increase
LVDQH[FHOOHQWTXDQWLWDWLYHLQGLFDWRURIPRUSKLQHZLWKGUDZDO
Nicotine-induced antinociception is reduced by opioid receptor
DQWDJRQLVWQDOR[RQHZKLOHQLFRWLQHLQGXFHG6&6LQFUHDVHLVQRWVXSSUHVVHGE\QDOR[RQHLQGLFDWLQJWKDWRSLRLGHUJLFV\VWHPPLJKW
participate in the nicotine-induced antinociception, but not SCS
increase. Nicotine-induced antinociception was reduced by both
D4E2 nAChR antagonist (dihydro-E-erythroidine; DH-E-E) andD7 nAChR antagonist (methyllycaconitine; MLA), while nicotineinduced SCS increase was suppressed by DH-E-E, but not MLA.
)XUWKHUPRUHQDOR[RQH PJNJ HOLFLWHG6&6HOHYDWLRQLQ
mice receiving repeated nicotine (3 mg/kg, twice a day for 7 days)
LQDGRVHGHSHQGHQWPDQQHUDQGWKHQDOR[RQHLQGXFHG6&6
increase was correlated with the doses (1-5 mg/kg, twice a day for
9 days) and the days (3 mg/kg, twice a day for 3-9 days) of repeated
QLFRWLQH:KHQRSLRLGDQWDJRQLVWQDOWUH[RQH ²PJNJ ZDV
administered together with repeated nicotine (5 mg/kg, s.c., twice
DGD\IRUGD\V QDOR[RQHLQGXFHG6&6LQFUHDVHZDVVLJQLÀFDQWO\suppressed, indicating that opioidergic system might participate in
the development of physical dependence on nicotine. Concomitantadministration of MLA with repeated nicotine, but not DH-E-E,
VXSSUHVVHGWKH6&6LQFUHDVHE\QDOR[RQH7KHVHUHVXOWVVXJJHVW
that opioidergic mechanism might be located on the downstreamof D7 nAChR, but not D4E2, in the central nervous system.

129(/.$33$23,2,'5(&(3725
$&7,9$7,1*6$/9,125,1$$1$/2*8(60206$/%$1'(206$/%$77(18$7('58*6((.,1*%(+$9,2856,17+(5$7
$1'02'8/$7('23$0,1(75$163257(5)81&7,21
Simonson, B.1, Morani, A.1, Ewald, A.1, Walker, L.1,
3ULVLQ]DQR72, *Kivell. B.1(1) School of Biological Sciences,Victoria University of Wellington, New
Zealand, (2) Department of Medicinal Chemistry, School of Pharmacy,
University of Kansas

0RQGD\²SP
Activation of kappa-opioid receptors (KOPr) by Salvinorin A (SalA) attenuates cocaine prime-induced drug-seeking in rats withfewer side effects compared to traditional arylacetamide KOPr
agonists. In the present study, we investigate the behaviouralwww.dcconferences.com.au/inrc2013/

DQWLDGGLFWLRQHIIHFWVDQGVLGHHIIHFWSURÀOHRIWZRSRWHQWORQJHUDFWLQJ6DO$DQDORJXHVPHWKR[\PHWK\O6DOYLQRULQ% 0206DO
% DQGHWKR[\PHWK\OVDOYLQRULQ% (206DO% DQGHYDOXDWHWKHLU
ability to modulate the function of the dopamine transporter(DAT). Our aim is to identify novel anti-addiction compoundswith reduced side effects and identify their mechanism of action.
%RWK0206DO%DQG(206DO%VLJQLÀFDQWO\DWWHQXDWHFRFDLQH
prime-induced drug-seeking behaviour in cocaine self-administeringrats at a dose of 0.3 mg/kg. However, increased immobility anddecreased swimming in the forced swim test indicates depressive
side effects similar to those seen in the parent compound, Sal A.
MOM Sal B rapidly increases DAT function in tissue taken from
WKHUDWVWULDWXPQXFOHXVDFFXPEHQVDQGPHGLDOSUHIURQWDOFRUWH[using rotating disk electrode voltametry techniques. The dopamineXSWDNHNLQHWLFVLQWKHQXFOHXVDFFXPEHQVVKRZHGDVLJQLÀFDQWLQFUHDVHLQ9PD[  ZLWKQRFKDQJHLQ.P  FRPSDUHGWRFRQWUROV 9PD[  DQG.P 
0.6) (p<0.05)), indicating an increase in the number of functional
WUDQVSRUWHUVZLWKQRFKDQJHLQGRSDPLQHELQGLQJDIÀQLW\7KH
effects on DAT function were reversed by pre-incubation with
KOPr antagonist nor-BNI. Both MOM Sal B (p<0.01) and EOM
Sal B (p<0.05) rapidly phosphorylate ERK in vitro in a KOPrGHSHQGHQWPDQQHUXVLQJSKRVSKRVSHFLÀFDQWLERGLHVDQG:HVWHUQ
blotting techniques. Inhibition of ERK activation by U0126, a MEK
inhibitor also prevented the KOPr mediated increases in DATfunction. This study shows two novel potent Sal A analogues thathave behavioural anti-addiction effects and modulate DAT function
in a KOPr and ERK dependent manner.
Supported by Health Research Council of New Zealand, Neurological Foundationof New Zealand, and Wellington Medical Research Foundation.

0,&52*/,$/$&7,9$7,2135(&('(6$17,
23,2,'6<67(0,10253+,1($1$/*(6,&
TOLERANCE
0DWVXVKLWD<DQG8HGD+
Dept of Mol Pharmacol & Neurosci, Nagasaki Univ Grad Sch of Biomed Sci,
Nagasaki

7XHVGD\²SP
It is well known that the analgesic tolerance and dependence
caused by chronic morphine treatment are associated withneural plasticity at the cellular, synaptic and neural circuit levelsin the central nervous system. However, little is known of theplasticity mechanisms in terms of neural circuit. Our previous
studies demonstrated that NMDA receptor and brain-derived
neurotrophic factor (BDNF) play key roles in the development ofmorphine analgesic tolerance. In addition to these mechanisms, weÀUVWO\UHSRUWHGWKDWPRUSKLQHLQGXFHVWKHDFWLYDWLRQRIFXOWXUHG
PLFURJOLDLQWHUPVRIPRUSKRORJLFDOFKDQJHDQGJHQHH[SUHVVLRQ
of BDNF, which is associated with NMDA. In the present study,
we attempted to clarify whether all these three components
are all involved, if they work, which precedes other machineries.
When mice are given with chronic morphine for 6 days, microglial
activation was only observed at the early stage. The injectionof minocycline to inhibit microglial activation or liposomeencapsulated chrodronate to disrupt activated microglia abolishedmorphine analgesic tolerance. Although, the up-regulation of
BDNF and NMDA receptor subunit, NR2A in western blot wereobserved in the PAG, which is the major brain region involved
in morphine analgesia, the treatment with minocycline and the
LQMHFWLRQRI%'1)DQWLERG\DEROLVKHG%'1)DQG15$H[SUHVVLRQin PAG. Thus, these results suggest that microglial activation,
BDNF release and NMDA receptor play pivotal roles in morphineanalgesic tolerance via anti-opioid machineries.

51

POSTER ABSTRACTS

())(&7,9(1(662)$0,',12
7$3$$*$,1670253+,1(5(6,67$17
1(8523$7+,&3$,1

92/817$5<$/&2+2/,17$.($1'&%
$1'0235(&(3725'(16,7<,1287%5('WISTAR RATS

*Mizoguchi, H.1, Watanabe, C.1<RQH]DZD$1, Sakurada,T.2,
Sakurada, S.1*Momeni, S., Bergström, L. and Roman, E.

(1) Department of Physiology and Anatomy,Tohoku Pharmaceutical
University, Sendai, Miyagi 981-8558, Japan(2) First Department of Pharmacology, Daiichi College of Pharmaceutical
Sciences, Fukuoka, Fukuoka 815-8511, Japan

0RQGD\²SP$PLGLQR7$3$LVDSHSWLGLFDQDOJHVLFZKLFKVKRZVKLJKDIÀQLW\and selectivity for PRSLRLGUHFHSWRUVDQGSURGXFHVH[WUHPHO\potent and longer lasting antinociception. Unlike traditional
narcotic analgesics, the spinal antinociception of amidino-TAPA
is mediated through the activation of distinct P-opioid receptors,
MOR-1J, MOR-1K and MOR-1L, which are amidino-TAPA-sensitive
but morphine-insensitive splice variants. Most notable characterof amidino-TAPA is its effectiveness against morphine-resistantneuropathic pain. In the present study, the antinociceptive
mechanism of amidino-TAPA against the neuropathic pain wasinvestigated.
The neuropathic pain model was developed in mice according to
WKHPHWKRGE\6HOW]HUDQGKLVFROOHDJXHVZLWKPLQRUPRGLÀFDWLRQ
The antinociceptive effects of morphine and amidino-TAPA
ZHUHPHDVXUHGXVLQJWKHYRQ)UH\ÀODPHQWDWGD\VDIWHUWKH
nerve ligation. As reported well, the spinal antinociception ofmorphine was markedly suppressed in ipsilateral paw in comparedin contralateral paw. In contrast, amidino-TAPA showed same
magnitude of spinal antinociception in both paws. In mice thePRSLRLGUHFHSWRUVSOLFHYDULDQWVFRQWDLQLQJH[RQRI025
gene had been knocked down, the spinal antinociception inducedby amidino-TAPA was attenuated in both paws with the same
magnitude. In contrast, the spinal antinociception induced byamidino-TAPA was more strongly attenuated in ipsilateral paw thanin contralateral paw under the condition that MOR-1J, MOR-1K
or MOR-1L had been knocked down. On the other hand, thespinal antinociception of morphine in sham-operated mouse was
DWWHQXDWHGXQGHUWKHFRQGLWLRQZKLFKH[RQFRQWDLQLQJVSOLFH
variants, but not MOR-1J, MOR-1K or MOR-1L, had been knocked
GRZQ$IWHUWKHVFLDWLFQHUYHOLJDWLRQP51$H[SUHVVLRQRIH[RQ
1-containing splice variants, but not MOR-1K or MOR-1L, weremarkedly reduced in DRG of ipsilateral side in compared in DRG
RIFRQWUDODWHUDOVLGH2QWKHFRQWUDU\P51$H[SUHVVLRQRIDQ\splice variants in spinal cord was not affected after the sciatic
nerve ligation.
In conclusion, the reduction of spinal antinociception of morphine
after sciatic nerve ligation may be caused by the reduction of themorphine-sensitive splice variants in DRG. On the contrary, thespinal antinociception of amidino-TAPA is maintained after thesciatic nerve ligation, since amidino-TAPA-sensitive splice variants
in DRG are maintained.

52

Neuropharmacology, Addiction and Behaviour, Department of Pharmaceutical
Bioscience, Uppsala University, Sweden

7XHVGD\²SP
Alcohol use disorders (AUD) are heterogeneous with regard
to etiology, liability for addiction and response to treatment.
,QGLYLGXDOYXOQHUDELOLW\LVGHSHQGHQWRQDFRPSOH[LQWHUDFWLRQ
between different neurotransmitter systems. The endocannabinoid
and opioid systems are implicated in AUD. Antagonists to the
CB1 receptor, e.g. rimonabant, as well as the MOP receptor,
HJQDOWUH[RQHGHFUHDVHVYROXQWDU\DOFRKROLQWDNHDQGDOFRKRO
consumption modulates receptor density and receptor-mediatedsignalling.
The aim of the present study was to investigate a possible
association between behavioural characteristics, voluntary alcohol
intake and alcohol-induced effects on CB1 and MOP receptor
density.
Forty adult male outbred Wistar rats were tested in the openÀHOGDQGWKHPXOWLYDULDWHFRQFHQWULFVTXDUHÀHOGWHVWVSULRUWR
alcohol access. The rats were randomly assigned into water or
DOFRKROJURXSV7KHDQLPDOVKDGDFFHVVWRHWKDQROVROXWLRQ
using a two-bottle binge-like free-choice paradigm, with 24 houraccess for three consecutive days per week for seven weeks. Ten toGD\VDIWHUWKHÀQDODOFRKROVHVVLRQWKHDQLPDOVZHUHGHFDSLWDWHGthe brains were immediately frozen and later sectioned into 12
mm coronal sections. The CB1 ligand [3H] SR141176 and the MOP
ligand [3H] DAMGO were used in an autoradiography method
followed by densiometric measurements.
During the last week of access to alcohol the average (± SEM)
ethanol intake was 3.3 ± 0.2 g/kg. The alcohol-drinking rats had
XSUHJXODWHG&%UHFHSWRUGHQVLW\LQFLQJXODWHDQGPRWRUFRUWH[and also up-regulated MOP receptors in the posteromedial cortical
amygdaloid nucleus relative to water-drinking rats. In the central
and medial amygdaloid nucleus, alcohol consumption resulted in
down-regulation of CB1 receptor density. MOP receptor density
was down-regulated in the globus pallidus, lateral septal nucleus
and the nucleus accumbens compared to the water-drinking
controls. These differences were revealed after the 11-day timeinterval between the last alcohol session and decapitation. The
results reveal long-lasting alterations in areas of relevance for risktaking behaviour, reinforcement and addiction processes.
Supported by the Alcohol Research Council of the Swedish Alcohol Retailing
Monopoly, and the Facias, the M Bergvall, the Å Wiberg and the F & I Thuring
Foundations.

International Narcotics Research Conference 2013

POSTER ABSTRACTS

,192/9(0(172)7+(/21*&+$,1)$77<$&,'5(&(3725*35,1&)$,1'8&(',1)/$00$725<3$,102'(/0,&(*Nakamoto, K.1, Nishinaka,T.1, Sato, N.1, Mankura, M.2,
.R\DPD<3, Kasuya, F.4 and Tokuyama, S.1
1
Department of Clinical Pharmacy, Kobe Gakuin University, School of
Pharmaceutical Sciences, Kobe, Japan. 2 Bizen Kasei Chemical Co. Ltd.,
Akaiwa city, Okayama, Japan, 3 Laboratory of Pharmacology, Faculty of
Pharmacy, Osaka Ohtani University, Osaka, Japan. (4) Biochemical Toxicology
Laboratory, Faculty of Pharmaceutical Sciences, Kobe Gakuin University, Kobe,
Japan.

0RQGD\²SP
GPR40 has been reported to be activated by long-chain fatty acids,
VXFKDVGRFRVDKH[DHQRLFDFLG '+$ +RZHYHUUHSRUWVVWXG\LQJ
functional role of GPR40 in the brain are lacking. The present study
focused on the relationship between pain regulation and GPR40,
investigating the functional roles of brain GPR40 during chronic
pain caused using a Complete Freund's Adjuvant (CFA)-inducedLQÁDPPDWRU\SDLQPRXVHPRGHO/RQJODVWLQJK\SHUSODVLDRISDZ
persistent mechanical allodynia and thermal hyperalgesia wereelicited in CFA-treated mice. Under these conditions, GPR40
SURWHLQH[SUHVVLRQLQWKHK\SRWKDODPXVDQGPHGXOODREORQJDWDZDV
increased at 7 days after CFA injection, in comparison with thesaline control group, but not at 1 or 3 days. GPR40 was colocalizedwith NeuN, a neuron marker, but not with GFAP, an astrocyte
PDUNHU*)$3SURWHLQH[SUHVVLRQZDVPDUNHGO\LQFUHDVHGLQthe hypothalamus at 1 day after CFA injection. This increase was
VLJQLÀFDQWO\LQKLELWHGE\WKHLQWUDFHUHEURYHQWULFXODULQMHFWLRQRI
ÁDYRSLULGRO QPRO DF\FOLQGHSHQGHQWNLQDVHLQKLELWRUGHSHQGLQJ
on the decreases in both the increment of GPR40 protein
H[SUHVVLRQDQGWKHLQGXFWLRQRIPHFKDQLFDODOORG\QLDDWGD\VDIWHU
CFA injection. The i.c.v. injection of DHA (50 Pg) and GW9508(1.0 PJ D*35VHOHFWLYHDJRQLVWVLJQLÀFDQWO\UHGXFHGPHFKDQLFDO
allodynia at 7 days after CFA injection. These effects were inhibitedby i.c.v. pretreatment with GW1100 (10 Pg), a GPR40 antagonist.
7KHVHÀQGLQJVVXJJHVWHGWKDWEUDLQ*35PLJKWKDYHKDGDQ
important role in this pain control system. Furthermore, astrocytes
might have contributed to the observed regulation of GPR40
SURWHLQH[SUHVVLRQ
6XSSRUWHGE\JUDQWVLQDLGIURPJUDQWVLQDLGIRU6FLHQWLÀF5HVHDUFK & (24592364) from the Ministry of Education, Culture, Sports, Science, andTechnology of Japan.

*3527(,1*$7(',1:$5'/<5(&7,)<,1*
327$66,80 *,5. &+$11(/63/$<$
35,0$5<52/(,17+($17,12&,&(37,9(())(&72)2;<&2'21(%871270253+,1($1')(17$1</$76835$63,1$/6,7(6*Ono, H.1, Nakamura, A.1, Fujita, M.1, Shibasaki, M.2,
Mori,T.2, Suzuki,T.2, Sakaguchi, G.1, Hasegawa, M.1 andKanemasa,T.1
1
Pain & Neurology, Medicinal Research Laboratories, SHIONOGI Co., Ltd.,
Osaka, Japan2
Department of Toxicology, Hoshi University School of Pharmacy andPharmaceutical Sciences,Tokyo, Japan

7XHVGD\²SP
2[\FRGRQHPRUSKLQHDQGIHQWDQ\ODUHPXRSLRLGUHFHSWRU 025 
agonists prescribed to control moderate to severe pain. Previous
VWXGLHVVXJJHVWHGWKDWWKHVHRSLRLGVH[KLELWGLIIHUHQWDQDOJHVLF
SURÀOHV:HWKHUHIRUHK\SRWKHVL]HGWKDWGLVWLQFWPHFKDQLVPV
mediate the differential effects of these opioids and investigated
the role of G protein-gated inwardly rectifying potassium (GIRK)
channels in their supra-spinal and spinal antinociceptive effects. The
DQWLQRFLFHSWLYHHIIHFWVRIR[\FRGRQHZLWKERWKLFY QPRO 
www.dcconferences.com.au/inrc2013/

and i.t. (1-30 nmol) administration were markedly and similarly
attenuated by tertiapin-Q (30 pmol, i.c.v. and i.t.), a selective GIRKFKDQQHOLQKLELWRULQPLFHWDLOÁLFNWHVW,QFRQWUDVWLFYPRUSKLQH
(0.1-3 nmol) and fentanyl (0.03-1.7 nmol) administration wasinsensitive to GIRK channel inhibition, while i.t. morphine (0.1-3nmol) and fentanyl (0.03-1.7 nmol) administration was markedly
DWWHQXDWHG8VLQJERQHFDQFHUSDLQPRGHOZHIXUWKHUH[DPLQHG
the antinociceptive effect of these opioids. Antinociceptive effect
RIVXEFXWDQHRXVDGPLQLVWUDWLRQRIR[\FRGRQH PJNJ EXWQRW
that of morphine (30 mg/kg) and fentanyl (0.1 mg/kg), was strongly
attenuated in the presence of tertiapin-Q (30 pmol, i.c.v.), indicating
the importance of GIRK channel function in the antinociceptive
effect in an opioid-dependent manner. These results demonstrated
that GIRK channels play a primary role in the antinociceptive
HIIHFWVRIR[\FRGRQHEXWQRWPRUSKLQHDQGIHQWDQ\ODWVXSUDVSLQDO
sites, and suggested that supraspinal GIRK channels are responsible
IRUWKHXQLTXHDQDOJHVLFSURÀOHRIR[\FRGRQH

+286,1*&21',7,216$))(&7%$6$/
$1'$/&2+2/,1'8&('23,2,'/(9(/6,1
ADOLESCENT RATS
*3DOP6DQGNylander, I.
Neuropharmacology, Addiction & Behaviour, Department of Pharmaceutical
Biosciences, Uppsala University, Uppsala, Sweden

0RQGD\²SP
The opioid system is involved in alcohol-induced reward andaddiction but our knowledge of alcohol-induced effects on opioids
before and after adolescent drinking is sparse. Many of the animalmodels of voluntary intake are based on single housing, but fora social animal such as a rat this is stressful, even more so inadolescent animals. In the current study we therefore address the
LVVXHRIKRXVLQJFRQGLWLRQVDJHDQGWLPHRIH[SRVXUHWRDOFRKRO
and their combined effects on opioid peptide levels in the rat brain.
7KUHHJURXSVRIUDWVZHUHXVHGDQGZHHNROGUDWVH[SRVHG
RQFHWRDOFRKRODQGDWKLUGJURXSRIZHHNROGUDWVH[SRVHG
WRDOFRKROIRUVL[ZHHNVLQDYROXQWDU\ELQJHGULQNLQJPRGHO
with sessions three times a week. All alcohol-drinking animals
were single housed. Control groups were age-matched water
drinkers that were single or group housed. Brain structures andthe pituitary gland were dissected and the immunoreactive levels
RIWKHRSLRLGSHSWLGHVG\QRUSKLQ% '<1% PHWHQNHSKDOLQ
Arg6Phe7 (MEAP) and beta-endorphin (BEND) were analysedusing radioimmunoassay.
1RWVXUSULVLQJO\H[WHQVLYHDJHGLIIHUHQFHVZHUHIRXQG+RXVLQJFRQGLWLRQVZHUHLPSRUWDQWIRU0($3DQG'<1%LQDOODJHV
but the structures affected differed between ages. Interactions
EHWZHHQKRXVLQJFRQGLWLRQDQGDJHZHUHIRXQGWRDIIHFW'<1%LQ
WKHPHGLDOSUHIURQWDOFRUWH[VWULDWXPDQGKLSSRFDPSXV6HYHUDO
alcohol-induced effects were found, but there were few differences
between long-term drinking and an acute session. However, MEAP
LQWKHK\SRWKDODPXVPHGLDOSUHIURQWDOFRUWH[DQGKLSSRFDPSXVUHVSRQGHGRSSRVLWHO\WRDFXWHRUORQJWHUPH[SRVXUH
The results show that housing conditions affect opioid levels to
DODUJHH[WHQWDQGWKDWWKHVHHIIHFWVFDQEHGLIIHUHQWGHSHQGLQJ
on age and duration. These effects can in turn affect thealcohol-induced changes. Housing conditions can therefore be
an important confounding factor, but is often neglected in theliterature. In light of the importance of social interactions duringadolescence, affordable models, in which both social conditions andalcohol-induced effects can be studied, need to be developed andevaluated.
Funded by SRA (SRA 2011-0056), the Swedish Medical Research Council (201261X-22090-01-3) and ERAB (EA 11 30).

53

POSTER ABSTRACTS

48.THE INTERACTION OF SEVERAL$17,'(25(66$17'58*6:,7+$&87($1'
&+521,&0(7+$'21,10,&($1'3266,%/(&/,1,&$/,03/,&$7,216
Schreiber, S.1, Hostovsky, A.2, %DUDN<2, Rubovitch,V. 2, and3LFN&*2*
Department of Psychiatry,Tel Aviv Sourasky Medical Center & Tel Aviv
8QLYHUVLW\6DFNOHU)DFXOW\RI0HGLFLQH2Department of Anatomy,Tel-Aviv
University Sackler Faculty of Medicine,Tel Aviv, Israel
1

7XHVGD\²SP
Background: Depression is highly prevalent among chronic-pain
patients and methadone maintenance treatment (MMT) patients.
Controversy regarding effective antidepressant treatment for thesepatients persists.
Objective: Assessing the two models in mice, using the hotplate
assay.
Methods: In the model of depressed pain patients, the impactof low, sub-threshold doses of 6 antidepressants with different
PHFKDQLVPVRIDFWLRQ ÁXYR[DPLQHHVFLWDORSUDPUHER[HWLQH
YHQODID[LQHGHVLSUDPLQHFORPLSUDPLQH RQWKHDQWLQRFLFHSWLYH
properties of a single (acute) dose of methadone. In the model ofdepressed MMT patients, the impact of low, sub-threshold dosesof 3 antidepressant drugs with different mechanisms of action(escitalopram, desipramine, clomipramine) on the antinociceptive
properties of chronic treatment with methadone.
Results: Following injection, acute methadone elicited analgesia
LQDGRVHGHSHQGHQWPDQQHU)OXYR[DPLQHDQGGHVLSUDPLQH
each at a sub-threshold dose induced a synergistic effect with
PHWKDGRQH(VFLWDORSUDPUHER[HWLQHYHQODID[LQHDQGFORPLSUDPLQH
given separately at a sub threshold dose induced no interaction.
Following two weeks of methadone administration (through animplanted mini-pump, i.p.) injection of escitalopram did not elicit
any analgesic effect, desipramine augmented the analgesic effect of
methadone, while clomipramine reduced it notably.
Conclusion: Possible clinical implications are that while
HVFLWDORSUDPUHER[HWLQHYHQODID[LQHDQGFORPLSUDPLQHGRQRW
affect acute methadone’s antinociception in mice and are safe to
EHJLYHQWRJHWKHUZLWKPHWKDGRQHÁXYR[DPLQHDQGGHVLSUDPLQH
notably augment methadone induced antinociceptive effects andshould be avoided due to the risk of inducing opiate-overdose.
Possible clinical implications of chronic methadone remain to be
determined as tricyclic antidepressants' cardiac effects may prevent
their use in MMT patients.

547+(3(5,$48('8&7$/*5$<&2175,%87(6722;<&2'21(%87127
0(7+$'21($17,12&,&(37,21,17+(5$7*Reid, R.A. and Morgan, M.M.,:DVKLQJWRQ6WDWH8QLYHUVLW\9DQFRXYHU:$86$

0RQGD\²SP
6WXGLHVH[DPLQLQJOLJDQGELDVHGVLJQDOLQJGHPRQVWUDWHWKDWRSLRLG
HIÀFDF\DQGWROHUDQFHFDQYDU\JUHDWO\GHSHQGLQJRQWKHRSLRLG
(Madia et al., 2009). Given that opioids such as morphine inhibit
pain, in part, by binding to mu-opioid (MOP) receptors in theperiaqueductal gray (PAG), microinjection of opioids directly into
the PAG provides an opportunity to link ligand-biased signaling at
the MOP receptor to antinociception. Previous research has shownthat administration of morphine, fentanyl, DAMGO, dermorphin,or ß-endorphin into the PAG produces antinociception (Bobeck
et al., 2009; Bodnar et al., 1988; Macey et al., 2010). Although it
is assumed that microinjection of any MOP receptor agonist
into the PAG would produce antinociception, this hypothesis
has not been tested. The present study tested this hypothesis bymeasuring antinociception and tolerance to PAG microinjection
RIWKHFRPPRQO\XVHG023UHFHSWRUDJRQLVWV·R[\FRGRQHDQG
methadone. Male Sprague-Dawley rats were implanted with aguide cannula aimed at the ventrolateral PAG and allowed 1 week
to recover before behavioral testing. Microinjection of ¼ log
FXPXODWLYHGRVHVRIR[\FRGRQHLQWRWKH3$*SURGXFHGDGRVH
dependent inhibition of nociception (ED50 = 191 µg) measured
with the hot plate test. Analysis of the time course for thisantinociception revealed a peak effect at 5 min and duration of
DSSUR[LPDWHO\PLQ7ZLFHGDLO\LQMHFWLRQVRIR[\FRGRQHLQWRWKH
3$*IRUGD\VGLGQRWDOWHUWKHR[\FRGRQHGRVHUHVSRQVHFXUYH
measured on Day 3, indicating a lack of tolerance to repeated
R[\FRGRQHLQMHFWLRQV,QFRQWUDVWWRR[\FRGRQHWKHUHZDVQR
dose (1.0, 5.6, 10, 56, 100 µg/ 0.4 µL) at which microinjection of
methadone into the PAG produced antinociception. These data
demonstrate that the antinociception produced by activation of
MOP receptors in the ventrolateral PAG are engaged by some,
but not all MOP receptor agonists. Moreover, previous studies
showing tolerance to morphine or fentanyl microinjections into
the ventrolateral PAG (Bobeck et al., 2012) are in direct contrast
WRR[\FRGRQHZKLFKGRHVQRWDSSHDUWRSURGXFHWROHUDQFHZLWK
repeated injections into the PAG.
This study was supported by NIH grant DA015498.

International Narcotics Research Conference 2013

POSTER ABSTRACTS

,1&5($6('5(*,21$/*5$<0$77(5
92/80(,123,2,'5(&(3725.12&.287
0,&($6'(7(50,1('%<05,92;(/%$6('0253+20(75<3+<6,2/2*,&$/$1'
3$7+23+<6,2/2*,&$/5(*8/$7,212),/
(;35(66,21%<12&,&(37,1253+$1,1)4
12)4 3(37,'(5(&(3725

Sasaki, K.a,b, Sumiyoshi, A.b, Nonaka, H.b, Hall, F.S.c8KO
G.R.c, Watanabe, M.d.DVDKDUD<a, Ikedae, K., Kawashima,R.b, Soraa, I.f

Donica, C.L.2=KDQJ<16LPSVRQ'XUDQG&'1, *Sherry,
D.M.2, 3, Gallucci, R.M.1, 2, Awwad, H.O.1, 2 and Standifer,
K.M.1, 2, 3

aDepartment of Biological Psychiatry,Tohoku University Graduate School of
Medicine, Sendai, JapanbDepartment of Functional Brain Imaging, Institute of Development, Aging
and Cancer,TohokuUniversity, Sendai, Japan
cMolecular Neurobiology Branch, NIDA/IRP/NIH, Baltimore, MD, USA
dDepartment of Anatomy, Hokkaido University Graduate School of Medicine,
Sapporo, JapaneAddictive Substance Project,Tokyo Metropolitan Institute of Medical Science,
Tokyo, JapanfDepartment of Psychiatry, Kobe University Graduate School of Medicine,
Kobe, Japan(1) Department Pharmaceutical Sciences, College of Pharmacy, (2)Oklahoma Center for Neuroscience and (3) Department Cell Biology, Collegeof Medicine, University of Oklahoma HSC, OKC, OK, USA

7XHVGD\²SPµ-opioid receptor knockout (MOR-KO) mice has revealed thecentral role of MORs in various behavioral functions including not
only analgesic effects but responses to stress, reward, tolerancecompared with wild type (WT) mice. However, it remains to
EHFRPSOHWHO\HOXFLGDWHGKRZRSLRLGUHFHSWRUVH[SUHVVHGLQ
neural circuits are related with particular behavioral functionsin these mice, and one possibility is that there reduced MOR
H[SUHVVLRQOHDGVWRFKDQJHVLQQHXUDOFLUFXLWU\,QWKLVVWXG\ZH
KDYHH[DPLQHGWKHEUDLQVRI025.2PLFHWRGHWHUPLQHWKH
volumetric changes associated with MOR deletion. We scanned
whole brain of 45 MOR-KO and 45 WT mice at 12 weeks of age
using high resolution, three-dimensional magnetic resonance T2
LPDJLQJ05,IRUDXWRPDWHGYR[HOEDVHGPRUSKRPHWU\ 9%0 
DQDO\VLV2XUÀQGLQJVZHUHDVIROORZV  025.2PLFHKDG
increased intracranial gray matter volume compared to WT
PLFH  VLJQLÀFDQWUHJLRQDOYROXPHWULFFKDQJHVZHUHGHWHFWHG
in periaqueductal gray (PAG), hypothalamus, olfactory bulb andcerebellum; (3) the greatest brain volume differences were
ORFDWHGDSSUR[LPDWHO\LQYHQWURODWHUDO3$*DUFXDWHQXFOHXVRI
hypothalamus, glomerular layer of olfactory bulb, copula of pyramis,
paramedian lobule of cerebellum, and cerebellar lobule. TheseÀQGLQJVVXJJHVWWKDWIDFWRUVRWKHUWKDQHOLPLQDWLRQRI025DORQH
may account for some of the behavioral differences observed in
WKHVHPLFHDQGWKDWOHYHOVRI025H[SUHVVLRQPD\LQÁXHQFHD
broader range of neural function in humans. Although the precise
relationship between volumetric changes and MOR deletion could
QRWEHGHWHUPLQHGEDVHGRQWKLVVWXG\DORQHRXUÀQGLQJVVXJJHVW
that morphological changes detected in MOR-KO mice by MRIVBM methods may relate to some of the behavioral differences
observed in MOR-KO mice.

www.dcconferences.com.au/inrc2013/

0RQGD\²SP
The neuropeptide, N/OFQ, has immunomodulatory actions in
DGGLWLRQWRPRGXODWLQJQRFLFHSWLRQDQ[LHW\DQGPDQ\RWKHUUHVSRQVHV([SUHVVLRQRI,QWHUOHXNLQ ,/ DF\WRNLQHOLQNHGWRLQÁDPPDWLRQDQGSDLQLVPRGXODWHGE\12)412)4SHSWLGH(NOP) receptor signaling at the mRNA and protein levels. To
better understand how N/OFQ modulates IL-6, IL-6 mRNA levels
in U937 human monocyte cells were assessed by quantitative
PCR (qPCR). Time course studies revealed that N/OFQ initially
increased IL-6 mRNA at 15 min, but decreased IL-6 mRNA over
WKHQH[WKU3UHWUHDWPHQWZLWKD123UHFHSWRUDQWDJRQLVW
blocked the N/OFQ-induced increase in IL-6 mRNA at 15 min,with no effect on the subsequent reduction in IL-6 at 4 hr. N/
OFQ-induced up-regulation of IL-6 mRNA also was blocked byinhibitors of NFNB and Akt signaling, but not by inhibitors of PKC,
p38 or ERK MAP kinase; suggesting potential mechanisms bywhich N/OFQ increases IL-6. We previously reported that Single
Prolonged Stress (SPS), a PTSD model, produced hyperalgesia andallodynia, and increased N/OFQ levels in serum, CSF and brain.
7RH[SORUHSRWHQWLDOUHODWLRQVKLSVEHWZHHQ12)4F\WRNLQHV
and PTSD-related pain, we assessed IL-6 mRNA and protein
levels in the spinal cord dorsal horn of male Sprague-Dawley rats
H[SRVHGWR636DQGPDWFKLQJFRQWUROVE\T3&5PXOWLSOH[F\WRNLQH
analysis and immunolabeling. SPS increased IL-6 mRNA in spinal
cord and brain between days 14-28 days, but IL-6 levels in sera
were unchanged. Preliminary studies indicate that treatment of
SPS rats with a NOP receptor antagonist from day 7-21 blocked
IL-6 protein up-regulation in spinal cord at day 21 and reversed
hyperalgesia and allodynia, suggesting that N/OFQ mediates IL-6
up-regulation during chronic stress and that this up-regulation may
contribute to allodynia and hyperalgesia in the PTSD model.
Animal protocol was approved by IACUC and the US Army Medical Research
DQG0DWHULHO&RPPDQG$QLPDO&DUHDQG8VH5HYLHZ2IÀFHFRPSOLHGZLWKWKH
Animal Welfare Act and adhered to the principles described in the Guide for Careand Use of Laboratory Animals.

55

POSTER ABSTRACTS

52. INVOLVEMENT OF P$1'G23,2,'
5(&(3725)81&7,21,17+(5(:$5',1*())(&72)  3(17$=2&,1(*Suzuki,T., Itoh,T., Saeki,T., Masukawa, D., Shibasaki, M.
and Mori,T.
Dept Toxicology, Hoshi University School of Pharmacy and Pharmaceuticalsciences,Tokyo 142-8501, Japan

7XHVGD\²SP
Most opioid receptor agonists have abuse potential, and therewarding effects of opioids can be reduced in the presenceof pain. While each of the enantiomers of pentazocine has a
GLIIHUHQWLDOSKDUPDFRORJLFDOSURÀOH  SHQWD]RFLQHKDVEHHQXVHG
clinically for the treatment of pain. However, little information isavailable regarding underlying mechanisms of rewarding effectsof (±)-pentazocine, and whether the (±)-pentazocine-induced
rewarding effects can be suppressed under pain. Therefore, thepresent study was performed to investigate the effects of painon the acquisition of the rewarding effects of (±)-pentazocine,
DQGWRH[DPLQHWKHPHFKDQLVPRIWKHUHZDUGLQJHIIHFWVRI
(±)-pentazocine using the conditioned place preference paradigm.
(±)-Pentazocine and (-)-pentazocine, but not (+)-pentazocine,
SURGXFHGVLJQLÀFDQWUHZDUGLQJHIIHFWV(YHQWKRXJKWKHUHZDUGLQJHIIHFWVLQGXFHGE\  SHQWD]RFLQHZHUHVLJQLÀFDQWO\VXSSUHVVHG
under pain induced by formalin, a high dose of (±)-pentazocine
SURGXFHGVLJQLÀFDQWUHZDUGLQJHIIHFWVXQGHUSDLQ,QWKHQRUPDO
condition, (±)-pentazocine-induced rewarding effects were
EORFNHGE\DORZGRVHRIQDOR[RQHZKHUHDVWKHUHZDUGLQJHIIHFWV
induced by high doses of pentazocine under pain were suppressedby naltrindole (a G-opioid receptor antagonist). Interestingly, SHQWD]RFLQHGLGQRWVLJQLÀFDQWO\DIIHFWGRSDPLQHOHYHOVLQWKH
QXFOHXVDFFXPEHQV7KHVHÀQGLQJVVXJJHVWWKDWWKHUHZDUGLQJ
effects of (-)-pentazocine may contribute to the abuse potential of(±)-pentazocine through P- as well as G-opioid receptors, which
depend on the pain situation, without robust activation of themesolimbic dopaminergic system.

1$/75(;21(2175($70(172)
1(8523$7+,&3$,1,10,&(/2&$//<
75$16)(&7(':,7+7+(087$170823,2,'
5(&(3725*(1(,163,1$/&25'
7DR3/1,3.DR-+1/DZ3<2 and Loh, H.H.2
Graduate Institute of Life Science, National Defence Medical Center,Taiwan,
52&2 Department of Pharmacology, University of Minnesota Medical
School, Minneapolis, MN, USA and 3 Center for Neuropsychiatric Research,
National Health Research Institutes,Taiwan, ROC
1

0RQGD\²SP
7KHRSLRLGDQWDJRQLVWVVXFKDVQDOR[RQHDQGQDOWUH[RQH
H[KLELWHGDJRQLVWLFSURSHUWLHVDWWKHPXWDWHGPXRSLRLGUHFHSWRU
MORS196ACSTA, in which the conserved Ser196, Thr327,
and Cys330 were mutated to Ala, Ala, and Ser, respectively. In
RXUSUHYLRXVVWXG\V\VWHPLFQDOR[RQH PJNJVF HOLFLWHG
DQWLQRFLFHSWLYHHIIHFW GHWHUPLQHGE\WDLOÁLFNWHVW ZLWKRXW
the induction of tolerance, dependence, and rewarding effectin mice 2 weeks after intrathecal administration of dsAAV2MORS196ACSTA-EGFP. In the present study, we further
investigated whether this antinociceptive paradigm could be
effective in mice with neuropathic pain. The spinal nerve ligation(SNL) surgery was performed on mice 3 or 4 weeks after
intrathecal injection of the lentivirus which carried the gene of
MORS196ACSTA-EGFP. The von Frey tests were used to detect
WKHDQWLDOORG\QLFHIIHFWVRIV\VWHPLFPRUSKLQHRUQDOWUH[RQH

56

EHIRUHDQGDIWHUJHQHWUDQVIHFWLRQ$IWHUGD\VRIQDOWUH[RQHRU
morphine treatment (10 mg/kg, s.c., q.d.) from day 1 or day 6 after
surgery, the natural withdrawal signs were counted at 22 and 46
hours after the last drug injection. Our data have shown that the
LSVLODWHUDOSDZZLWKGUDZDOSUHVVXUHZDVVLJQLÀFDQWO\GHFUHDVHG
one day after SNL surgery and persisted at least for 19 days.
1DOWUH[RQH PJNJVF RUPRUSKLQH PJNJVF HOLFLWHG
VLJQLÀFDQWDQWLDOORG\QLDHIIHFWV7KHSDZZLWKGUDZDOSUHVVXUHDIWHUQDOWUH[RQHRUPRUSKLQHWUHDWPHQWZDVVLJQLÀFDQWO\LQFUHDVHG
when compared to saline treatment. The SNL-induced allodynia
was improved gradually and almost back to normal after chronic
QDOWUH[RQHRUPRUSKLQHWUHDWPHQWRQGD\+RZHYHUFKURQLF
WUHDWPHQWRIPRUSKLQH PJNJVFTG EXWQRWQDOWUH[RQH(10 mg/kg, s.c., q.d.) induced natural withdrawal signs. These data
LPSO\WKDWV\VWHPLFLQMHFWLRQRIQDOWUH[RQHPD\KDYHWKHUDSHXWLF
potential for chronic neuropathic pain without the development
RIGHSHQGHQFHDIWHUWKHJHQHWKHUDS\ZLWKWKHH[SUHVVLRQRIMORS196ACSTA in the spinal cord. (Supported by a grant from
National Science Council, Taiwan, ROC).

(67$%/,6+0(172)$&(175$/3267
6752.(3$,102'(/86,1**/2%$/
CEREBRAL ISCHAEMIC MICE
*Tokuyama, S.1, Harada, S.1, Nakamoto, K.1
1
Department of Clinical Pharmacy, Kobe Gakuin University, School of
Pharmaceutical Sciences, Kobe, Japan

7XHVGD\²SP
Stroke is the leading cause of disability in the world. Central poststroke pain (CPSP), an intractable secondary disease, is a serious
problem that occurs following cerebral stroke. However, thedetailed mechanisms underlying CPSP and standard treatments for
LWDUHQRWZHOOHVWDEOLVKHG7KHUHIRUHZHH[DPLQHGWKHQRFLFHSWLYH
threshold and alterations in the current stimulus threshold ofprimary afferent neurons in bilateral carotid artery occlusion(BCAO) and middle cerebral artery occlusion (MCAO) mice. Male
GG<PLFHZHUHVXEMHFWHGWRPLQRI%&$2RUKRI0&$27KH
development of mechanical and thermal hyperalgesia and changes
in current stimulus threshold in the hind paws were measured
after BCAO or MCAO using the von Frey test, plantar test anda Neurometer, respectively. The threshold for mechanical and
WKHUPDOK\SHUDOJHVLDLQERWKKLQGSDZVZDVVLJQLÀFDQWO\GHFUHDVHG
on day 3 after BCAO as compared with pre-BCAO treatment.
Furthermore, the sensitivity of C and AEÀEHUV DWVWLPXODWLRQRI
and 2000 Hz, respectively) was increased on day 3 after BCAO as
compared with pre-BCAO treatment, while that of AGÀEHUVZDV
not altered. On the other hand, in MCAO mice, the sensitivity of
AG and AEÀEHUV DWDQG+]VWLPXODWLRQUHVSHFWLYHO\ 
ZDVVLJQLÀFDQWO\LQFUHDVHGRQGD\DIWHU0&$2ZKLOHWKDWRI&ÀEHUV DW+]VWLPXODWLRQ ZDVXQDOWHUHG,QDGGLWLRQWKHSDZ
withdrawal threshold of the left hind paw as measured by the von)UH\ÀODPHQWWHVWZDVVLJQLÀFDQWO\GHFUHDVHGRQGD\DQGGD\after MCAO when compared with day 0, while that in the righthind paw was not different. Our data show the development ofbilateral hyperalgesia in these both models. Potentially, in BCAO
model, C and AEÀEHUVSHFLÀFK\SHUVHQVLWL]DWLRQDIWHUVWURNH
may have contributed to these symptoms. In MCAO model,
P\HOLQDWHGDÀEHUVSHFLÀFK\SHUVHQVLWL]DWLRQDIWHUVWURNHPD\KDYH
contributed to these symptoms.
6XSSRUWHGE\JUDQWVLQDLGIURPJUDQWVLQDLGIRU6FLHQWLÀF5HVHDUFK & (24592364) from the Ministry of Education, Culture, Sports, Science, andTechnology of Japan.

International Narcotics Research Conference 2013

POSTER ABSTRACTS

(*)31230,&(Toll, L.1, Wu, J.1, Mercatelli, D.1, Kieffer, B.2
1
2

Torrey Pines Institute for Molecular Studies, Port St. Lucie, FL, USA
Institut de Génétique et de Biologie Moléculaire et Cellulaire, Ilkirch, France

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The NOP receptor is the fourth member of the opioid receptor
family, and the cognate receptor for nociceptin/orphanin FQ, now
called N/OFQ. NOP receptors and N/OFQ are found throughout
the brain, in the spinal cord, dorsal root ganglia, and a variety of
peripheral tissues. To better characterize the localization andinternalization properties of the NOP receptor, we created mice
where the NOP receptor is replaced by an active EGFP-NOP
receptor fusion protein, similar to what has been previously
reported for an EGFP-delta opioid receptor. Binding studies with
[3H]N/OFQ have determined that homozygous and heterozygous
EFGP-NOP receptor containing mice have a similar receptor
number in brain to wild type litter mates, and that N/OFQ and avariety of selective and non-selective ligands have similar bindingDIÀQLWLHVWRWKHPXWDQWDQGZWUHFHSWRU(*)3123UHFHSWRU
are functional in the knock-in mice, as determined by [35S]GTPJS
binding, with NOP agonists having similar potency in the knockin and w.t. mice. The anatomic distribution of the EGFP-NOP
receptor was studied by immunohistochemical localization using
EGFP antibodies. Receptor localization was similar to what has
been reported from in vitro autoradiographic studies. Studies
H[DPLQLQJ12)4PHGLDWHGUHFHSWRULQWHUQDOL]DWLRQLQSULPDU\hippocampal cultures are ongoing. Direct visualization of the
NOP receptor will be a useful tool in understanding receptor
ORFDOL]DWLRQUHFHSWRUWUDIÀFNLQJDQGWKHFRORFDOL]DWLRQRI123
and mu opioid receptors.

6<1(5*,60%(7:((17+('(/7$23,2,'
$*21,6761&$1'$03+(7$0,1(2&&856
9,$$*/87$0$7(5*,&10'$5(&(3725
'(3(1'(170(&+$1,60
Bosse, K.E.1, Jutkiewicz, E.M.1, Mabrouk, O.S.2, Schultz,
K.N.2, Kennedy, R.T.2, Gnegy, M.E.1 and *Traynor, J.R.1
1
Department of Pharmacology and (2) Department of Chemistry, Universityof Michigan Ann Arbor, MI, 48109, USA.

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Delta opioid receptor (DOR) agonists have stimulant-like
properties. Thus, DOR agonists increase locomotor activity,
produce conditioned place preference in rodents, and generalize
to the discriminative stimulus effects of stimulants in rats andmonkeys. Despite these properties, the nonpeptidic DOR agonist
SNC80 is not self-administered. Furthermore, SNC80 fails to
SURPRWHGRSDPLQHHIÁX[IURPUDWVWULDWXPDQGGRHVQRWLQFUHDVH
H[WUDFHOOXODUGRSDPLQHOHYHOVLQWKHFDXGDWHSXWDPHQRUQXFOHXV
accumbens following systemic administration. Thus, SNC80 likely
modulates the dopaminergic system indirectly through othermechanisms or signaling pathways. Glutamate is known to cause
the release of dopamine through a Ca2+-sensitive mechanism thatinvolves activation of N-methyl-D-aspartate (NMDA) receptors.
+HUHZHH[DPLQHGWKHK\SRWKHVLVWKDW61&DFWVRQWKH
glutamatergic system to enhance both amphetamine-stimulated
GRSDPLQHHIÁX[DQGDPSKHWDPLQHVWLPXODWHGORFRPRWRUDFWLYLW\,QUDWVWULDWDOVOLFHV61&LQFUHDVHGH[WUDFHOOXODUJOXWDPDWHOHYHOVand decreased GABA levels. Inhibition of NMDA signaling withthe selective antagonist MK801 blocked the SNC80-mediatedwww.dcconferences.com.au/inrc2013/

HQKDQFHPHQWRIERWKDPSKHWDPLQHLQGXFHGGRSDPLQHHIÁX[DQG
ORFRPRWRUDFWLYLW\$GGLWLRQRIH[RJHQRXVJOXWDPDWHSRWHQWLDWHG
DPSKHWDPLQHVWLPXODWHGGRSDPLQHHIÁX[LQD0JDQG0.
sensitive manner. In the absence of Mg2+ SNC80 alone was
DEOHWRFDXVHGRSDPLQHHIÁX[DQGDOVRSURGXFHGDJUHDWHUHQKDQFHPHQWRIDPSKHWDPLQHHYRNHGGRSDPLQHHIÁX[6LQFHWKH
majority of DORs are located on inhibitory GABA terminals, wehypothesized that DOR activation inhibits GABA release, thus
GLVLQKLELWLQJJOXWDPDWHHIÁX[WRXOWLPDWHO\SURPRWHDPSKHWDPLQH
stimulated dopamine release. Perfusion of rat striatal slices with
WKH*$%$%UHFHSWRUDQWDJRQLVWK\GUR[\VDFORIHQLQFUHDVHGDPSKHWDPLQHVWLPXODWHGGRSDPLQHHIÁX[WRDVLPLODUH[WHQWDV
SNC80. Overall the results suggest that in the striatum, DOR
activation by SNC80 enhances glutamate neurotransmission, most
likely through GABA disinhibition, to regulate amphetamineVWLPXODWHGGRSDPLQHHIÁX[DQGDPSKHWDPLQHLQGXFHGEHKDYLRUV
primarily through activation of NMDA receptors. Supported by
R01 MH083754, R01 DA11697 and F31 DA019728.

,192/9(0(172)6835$63,1$/$1'
3(5,3+(5$/1$/2;21$=,1(,16(16,7,9(23,2,'5(&(37256,7(6,17+((;35(66,21
OF P23,2,'5(&(3725$*21,67,1'8&('
3+<6,&$/'(3(1'(1&(*8]DZD1Mori,T., Sugiyama, K., 6DLWRX<Shibasaki, M.,
Suzuki,T
Dept Toxicology, Hoshi University School of Pharmacy and Pharmaceuticalsciences,Tokyo 142-8501, Japan

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Withdrawal signs after the cessation of P-opioid receptor agonists
remains an obstacle in the clinical treatment of pain. There islimited information available on the mechanisms that underlie
WKHH[SUHVVLRQRIWKHZLWKGUDZDOVLJQVRIRSLRLGVDQGHVSHFLDOO\regarding the involvement of P-opioid receptor subtypes andthe location of the responsible opioid receptors. Therefore, thepresent study was designed to determine the mechanism of the
H[SUHVVLRQRIZLWKGUDZDOVLQJVLQP-opioid receptor agonistGHSHQGHQWPLFH0RUSKLQHR[\FRGRQHDQGIHQWDQ\OGHSHQGHQW
mice showed a marked loss of body-weight and other withdrawal
VLJQVDIWHUDQDOR[RQHFKDOOHQJH,QWHUHVWLQJO\WKHSKHQRW\SHRIWKHZLWKGUDZDOVLJQVIRUPRUSKLQHDQGR[\FRGRQHZDVGLIIHUHQWIURPWKDWRIIHQWDQ\O)XUWKHUPRUHSUHWUHDWPHQWZLWKQDOR[RQD]LQHGLG
QRWVLJQLÀFDQWO\DOWHUWKHZLWKGUDZDOVLJQVSUHFLSLWDWHGE\QDOR[RQH
in these P-opioid receptor agonist-dependent mice, whereas
WKHSHULSKHUDOO\OLPLWHGRSLRLGUHFHSWRUDQWDJRQLVWQDOR[RQH
PHWKLRGLGHVLJQLÀFDQWO\LQFUHDVHGWKHORVVRIERG\ZHLJKWDFFRPSDQLHGE\GLDUUKHDLQGLFDWLQJWKDWDSHULSKHUDOQDOR[RQD]LQH
insensitive site for opioid receptors, as an adaptation mechanism,
SOD\VDQLPSRUWDQWUROHLQWKHH[SUHVVLRQRIDWOHDVWWKHORVVRIERG\ZHLJKW2QWKHRWKHUKDQGLFYWUHDWPHQWZLWKQDOR[RQH
methiodide potently induced a jumping behavior and tremblingin morphine-dependent mice. These results indicate that the
prolonged activation of supraspinal P-opioid receptors plays a rolein most of the physical dependence induced by P-opioid receptor
agonists in mice. Thus, the withdrawal signs observed after the
cessation of P-opioid receptor agonists are distinctly regulated
WKRXJKVXSUDVSLQDODQGSHULSKHUDOQDOR[RQD]LQHLQVHQVLWLYHVLWHVRI
P-opioid receptors.

57

POSTER ABSTRACTS6(/)$'0,1,675$7,212)2;<&2'21(%<$'2/(6&(17$1'$'8/70,&(',))(5(17,$//<$))(&7675,$7$/
1(85275$160,77(55(&(3725*(1((;35(66,21

59. 7+(())(&72)23,2,'5(&(3725

=KDQJ<1, Schlussman, S.D.1, Butelman, E.R.1, Ho, A.1, Ott,
J.2, Kreek, M.J.1

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The Laboratory of the Biology of Addictive Diseases,The Rockefeller
8QLYHUVLW\1HZ<RUN1<,QVWLWXWHRI3V\FKRORJ\&KLQHVH$FDGHP\of Sciences, Beijing

1

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Rationale: Illicit use of prescription opioid analgesics (e.g.,
R[\FRGRQH LQDGROHVFHQFHLVDSUHVVLQJSXEOLFKHDOWKLVVXH2XU
JRDOZDVWRGHWHUPLQHZKHWKHUR[\FRGRQHVHOIDGPLQLVWUDWLRQ
differentially affects striatal neurotransmitter receptor gene
H[SUHVVLRQLQWKHGRUVDOVWULDWXPRIDGROHVFHQWFRPSDUHGWRDGXOWC57BL/6J mice.
Methods: Groups of adolescent mice (4 weeks old, n=12) and of
adult mice (11 weeks old, n=11) underwent surgery during which acatheter was implanted into their jugular veins. Mice were placed
LQVHOIDGPLQLVWUDWLRQFKDPEHUVIRUDFTXLVLWLRQRIR[\FRGRQHVHOI
administration or were yoked saline control after recovering fromVXUJHU\0LFHVHOIDGPLQLVWHUHGR[\FRGRQH PJNJLQIXVLRQ 
KUVGD\IRUFRQVHFXWLYHGD\V0LFHZHUHVDFULÀFHGZLWKLQone hour after the last self-administration session and the dorsal
VWULDWXPZDVLVRODWHGIRUP51$DQDO\VLV*HQHH[SUHVVLRQZDV
analyzed with real time PCR using a commercially available PCR
array.
Results: We found that adolescent mice self-administered less
R[\FRGRQHWKDQDGXOWPLFHRYHUWKHGD\V$GROHVFHQWVDQG
DGXOWVGLIIHUHGLQPRQRDPLQHR[\GDVH$ 0DRD DQGQHXURSHSWLGH<UHFHSWRUP51$LQWKHDEVHQFHRIR[\FRGRQHH[SRVXUHGD\R[\FRGRQHVHOIDGPLQLVWUDWLRQLQFUHDVHG0DRDP51$OHYHOV
compared to controls in both adults and adolescents. Gastrin
UHOHDVLQJSHSWLGHUHFHSWRUP51$VKRZHGDVLJQLÀFDQW'UXJ[Age interaction. More genes in the dorsal striatum of adolescentsFKDQJHGLQUHVSRQVHWRR[\FRGRQHVHOIDGPLQLVWUDWLRQFRPSDUHG
to controls than in adult mice.
Summary: 7KLVVWXG\GHPRQVWUDWHVIRUWKHÀUVWWLPHWKDWUHSHDWHGR[\FRGRQHVHOIDGPLQLVWUDWLRQGLIIHUHQWLDOO\DOWHUHG
QHXURWUDQVPLWWHUUHFHSWRUVJHQHH[SUHVVLRQLQWKHGRUVDOVWULDWXP
of adolescent versus adult mice.
Support: This work was supported by NIH-NIDA5'$$ <= DQG1,+1,'$'$ 0-.

58

32/<0253+,606215(&(3725)81&7,21*Knapman, A., Connor, M.
The Australian School of Advanced Medicine, Macquarie University, Sydney,
NSW, Australia

7KHUHLVVLJQLÀFDQWYDULDWLRQLQLQGLYLGXDOUHVSRQVHWRRSLRLG
drugs, one cause of which is likely to be polymorphisms on the
opioid receptors themselves. The µ-opioid receptor (MOR) is
the primary site of action for most analgesic opioids. Previous
VWXGLHVKDYHLGHQWLÀHGDQXPEHURIQDWXUDOO\RFFXUULQJVLQJOH
nucleotide polymorphisms (SNPs) in the coding region of the
MOR. The A118G SNP variant, present in various populations
DWDOOHOLFIUHTXHQFLHVUDQJLQJIURP²KDVEHHQDVVRFLDWHG
with differences in the requirement for post-operative opioid
analgesics, and for drug dependence, although these results are not
FRQVLVWHQW,QYLWUR025$*KDVEHHQUHSRUWHGWRH[KLELWDQ
DOWHUHGVLJQDOOLQJSURÀOHFRPSDUHGZLWK025LQFOXGLQJGLIIHUHQWȕ-endorphin potency and alterations in N-type Ca2+ channel
LQKLELWLRQKRZHYHUWKHVHÀQGLQJVDUHDOVRQRWFRQVLVWHQW)HZVWXGLHVKDYHH[DPLQHGWKHHIIHFWRI025SRO\PRUSKLVPVRQ
receptor function, or potential ligand functional selectivity. We
DVVHVVHGWKHUHODWLYHSRWHQF\DQGHIÀFDF\RIDUDQJHRIRSLRLG
ligands in Chinese hamster ovary (CHO) cells stably transfected
ZLWKKXPDQZLOGW\SH025DQGÀYHQDWXUDOO\RFFXUULQJ025
YDULDQWVLQFOXGLQJ$*5HFHSWRUH[SUHVVLRQOHYHOVZHUHVLPLODUIRUDOOPXWDQWVH[DPLQHG025PHGLDWHGDGHQ\O\OF\FODVH
$& LQKLELWLRQZDVPHDVXUHGXVLQJDQRYHOÁXRUHVFHQFHEDVHG
membrane potential assay. Treatment of CHO cells with the AC
activator forskolin (FSK) hyperpolarized CHO-MOR cells with aS(&RIWRDPD[LPXPRIIURPEDVHOLQH7KH
hyperpolarization induced by FSK (300nM) was inhibited in a dosedependent manner by the addition of a range of MOR agonists,
including DAMGO, morphine, ȕ-endorphin and buprenorphine (n ,Q025$*FHOOVWKHPD[LPDOEXSUHQRUSKLQHLQKLELWLRQ
RI$&ZDVUHGXFHGWR FRPSDUHGZLWKLQ025
WT, P < 0.05), without a change in potency, while responses to
PRUSKLQHDQG%HQGZHUHXQDIIHFWHG&HOOVH[SUHVVLQJ025&7VKRZHGVLJQLÀFDQWO\UHGXFHG(PD[DQGS(&IRUPRUSKLQH
but not DAMGO. By contrast, MOR-C253A cells showed
VLJQLÀFDQWO\LQFUHDVHG(PD[IRUPRVWOLJDQGVZLWKQRFKDQJHLQ
SRWHQF\&+2025&7ZDVFRQÀUPHGDVQRQIXQFWLRQDO7KH
differences in inhibition of AC observed between MOR variants
suggest that MOR SNPs may contribute to individual variability in
WKHUHVSRQVHWRRSLRLGDQDOJHVLFVDQGZHDUHH[SORULQJWKLVDWDrange of signalling pathways.

International Narcotics Research Conference 2013

POSTER ABSTRACTS62&,$/675(66(1*$*(623,2,'
02'8/$7,212)7+(/2&86&2(58/(86
125(3,1(3+5,1(6<67(0$1',1&5($6(6THE SALIENCE OF REWARD.
*Chaijale, N., Curtis A.L., Snyder, K., Bhatnagar,
S.,Valentino, R.J.
Childrens Hospital of Philadelphia, Philadelphia, PA. USA.

7XHVGD\²SP
The locus coeruleus-norepinephrine (LC-NE) system is amajor stress response system through which stress affects
arousal and cognitive function. The LC is co-regulated by stressneuromediators and endogenous opioids. Because social stress(SS) is a common stressor for humans, this study characterized
WKHHQGXULQJLPSDFWRIUHSHDWHG66RQ/&DFWLYLW\6SHFLÀFDOO\ZH
determined whether SS alters the LC-NE system two (Day 7) andten days (Day 15) after the last SS and evaluated whether thesechanges in LC-NE system translate to changes in cognitive function.
Rats were implanted with a multiwire bundle into the LC andH[SRVHGWRÀYHVHVVLRQVRIWKHUHVLGHQWLQWUXGHUPRGHORIVRFLDO
defeat. On Day 7, LC discharge rate was decreased in stressed rats
compared to controls (Day 1: 2.21±0.1Hz, Day 7: 1.35±0.1Hz forstress rats and Day 1: 2.23±0.1Hz, Day 7: 2.20±0.1Hz, for control
rats; n<40). By Day 15, LC rates were comparable between groups.
6\VWHPLFDGPLQLVWUDWLRQRIWKHRSLDWHDQWDJRQLVWQDOR[RQHUREXVWO\increased LC rate selectively in stressed rats when administered
RQ'D\ +]SUHQDOR[RQH+]SRVWQDOR[RQH 
RU'D\ +]SUHQDOR[RQH+]SRVWQDOR[RQH 
This cellular evidence of opiate withdrawal was accompanied bybehavioral signs of mild opiate withdrawal. To evaluate whether
66LQGXFHGFKDQJHVLQ/&DFWLYLW\DIIHFWFRJQLWLYHÁH[LELOLW\UDWV
were tested in an attention set shifting task (AST) and LC activity
was recorded. Stress rats performed better in intradimensional
set shifting (IDS): 36.7±4 and 23.8±4 trials to criterion for control
and stress rats, respectively (p<0.05). Recordings during IDS
showed that LC neurons of stressed rats were reward, but notdecision responsive. During reversal learning although overall
performance was comparable between groups, stressed rats madeless perseverative errors. LC neurons of stressed rats selectively
showed decision- and reward-related activation. LC activity also
increased for stressed rats only during incorrect trials between
decision and recognition of the absence of reward. These results
suggest that SS engages endogenous opioid modulation of LC
activity and induces a state of opiate dependence. Additionally, SS
strengthens the relationship between LC activity and goal-directedbehavior. Together these effects of SS on the LC system canincrease vulnerability to opiate abuse by promoting the positive
and negative reinforcing effects. Supported by T32-NS007413,
58077 LSDRP, MH40008, DA09082.

www.dcconferences.com.au/inrc2013/0253+,1(02'8/$7(6%5($67&$1&(5&(//0(7$67$7,&327(17,$/
Imani, B.A.1, Baran, J.2&DERW3-1 3DUDW01,2
12School of Pharmacy, University of Queensland,Woolloongabba, Australia
Department of Anesthesia Research, Cleveland Clinic, Cleveland, OH

Appropriate pain management during and after cancer surgery
may play a role in prevention of tumor recurrence and metastasis.
Opioids are proven to be highly effective perioperative analgesics
and are widely used in cancer surgery patients. Using a mousesyngeneic model of breast cancer, we studied the effect ofmorphine on tumor growth and dissemination to lungs. I.P.
injection of morphine (10mg/kg) to mice (n=8) every 12h for
3 consecutive days, caused a reduction in breast tumor growth
and tumor cell dissemination to the lungs as measured 18 days
after tumor inoculation. Morphine treatment also caused a
UHGXFWLRQLQFLUFXODWLQJSURWHRO\WLFHQ]\PHVRIH[WUDFHOOXODUPDWUL[(&0 PDWUL[PHWDOORSURWHLQDVH 003 DQGXURNLQDVHOLNH
plasminogen activator (uPA) measured by gelatin and caseinplasminogen zymography, respectively. Furthermore, we tested
the effect of morphine on co-cultures of breast cancer cellswith stromal cells, either endothelial cells or macrophages. In
co-cultures of 4T1 breast cancer cells with murine RAW264.7
PDFURSKDJHVRUPXULQH+9HQGRWKHOLDOFHOOVWKHOHYHORIPDWUL[proteases produced by cells was increased, and so was the level
RIPDWUL[SURWHDVHLQKLELWRUV7,03V,QWHUHVWLQJO\PRUSKLQH
treatment of these co-cultures reduced the level of MMP-9 andincreased its endogenous inhibitor, TIMP-1, thereby altering theRYHUDOOSURWHRO\WLFSURÀOH0RUSKLQHDIIHFWHGFRFXOWXUHVEXW
not cells grown individually. This suggests that anti-tumor effects
of morphine are mediated through modulation of paracrine
FRPPXQLFDWLRQEHWZHHQFDQFHUFHOOVDQGLQÀOWUDWLQJFHOOV

59

DELEGATE
LIST$3URI0LFKDHO0F'RQRXJK
Western Health Australia

Dr Sulie Lin Chang
Seton Hall University United States

mikemcdo@bigpond.net.ausulie.chang@shu.edu

Dr Andrew Alt
Bristol-Myers Squibb United States'U<XORQJ&KHQ
Binghamton University United States

andrew.alt@gmail.com

ylchen@binghamton.edu

0U<XWD$RNL7RKRNXPharmaceutical University Japan

3URI/LK&KX&KLRX
Department of Pharmacology, College of Medicine, National Taiwan
University Taiwan

ay055002@yahoo.co.jp

Dr Derrick Arnelle
Alkermes United States
derrick.arnelle@alkermes.com

Dr Seksiri Arttamangkul
Vollum Institute / Oregon Health & Science University United Statesarttaman@ohsu.edu

Dr Brady Atwood
National Institute on Alcohol Abuse and Alcoholism United States
brady.atwood@nih.gov'U$QQD0DULH%DEH\
University of New England Australia
annamarie.babey@une.edu.au

Dr Elena Bagley
Discipline of Pharmacology, University of Sydney Australia
elena.bagley@sydney.edu.au

3URI%HUQDUG%DOOHLQH
University of Sydney Australia
bernard.balleine@sydney.edu.au

3URI/DQ%DR
Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences
Chinabaolan@sibcb.ac.cn

3URI6HOHQD%DUWOHWW
Queensland University of Technology Australia
selena.bartlett@qut.edu.au

Dr Helene Beaudry
Columbia University United States
helene.beaudry@usherbrooke.ca

Dr Mei Bigliardi
Clinical Research Unit for Skin Allergy Regeneration Singapore
mei.bigliardi-qi@imb.a-star.edu.sg'U3DXO%LJOLDUGLAgency for Science,Technology & Research Singapore
paul.bigliardi@imb.a-star.edu.sg

Dr William Birdsong
Oregon Health & Science University United Statesbirdsong@ohsu.edu

Dr Stephanie Borgland
University of Calgary Canadas.borgland@ucalgary.ca

Dr Meritxell Canals
Monash Institute of Pharmaceutical Sciences Australia
meri.canals@monash.edu

Dr Nayla ChaijaleChildren's Hospital of Philadelphia United States
naylachaijale@hotmail.com

lcchiou@ntu.edu.tw

3URI0DF&KULVWLH
University of Sydney Australia
mac.christie@sydney.edu.au

3URI$UWKXU&KULVWRSRXORVMonash Institute of Pharmaceutical Sciences, Monash University
Australia
arthur.christopoulos@monash.edu

3URI6DQGUD&RPHU
Columbia University United States
sdc10@columbia.edu

3URI0DUN&RQQRU
Australian School of Advanced Medicine Australia
mark.connor@mq.edu.au

Dr Alistair Corbett
Glasgow Caledonian University United Kingdom
A.d.corbett@gcu.ac.uk

Dr Brian Cox
Uniformed Services University United States
EULDQFR[#XVXKVHGX

Mrs Loudin Daoura
Uppsala University Sweden
Loudin.Daoura@farmbio.uu.se

0U=ROWDQ'HNDQInstitute for Molecular Bioscience Australia
z.dekan@imb.uq.edu.au

3URI/DNVKPL'HYL
Lakshmi Devi United States
lakshmi.devi@mssm.edu

Dr Nikolas Dietis
Tasmanian School of Pharmacy Australia
Nikolas.Dietis@utas.edu.au

3URI.DWH'RODQUniversity of NSW Australia
k.dolan@unsw.edu.au

Dr Adrian Dunlop
Hunter New England Local Health District Australia
adrian.dunlop@hnehealth.nsw.gov.au

Dr Wakako Fujita
Icahn School of Medicine at Mount Sinai United States
wakako.fujita@mssm.edu

Ms Maja Galic
Grunenthal Australia Australia
Maja.Galic@grunenthal.com

3URI/RXLV*HQGURQUniversity De Sherbrooke Canadalouis.gendron@usherbrooke.ca

60International Narcotics Research Conference 2013

DELEGATE LIST

'U=DÀURXOD*HRUJRXVVL1DWLRQDO&HQWUHIRU6FLHQWLÀF5HVHDUFK*UHHFH
iro@bio.demokritos.gr

Dr Robert Gereau
Washington University School of Medicine United States
gereaur@wustl.edu

Miss Linnea Granholm
Uppsala University Sweden
linnea.granholm@farmbio.uu.se

3URI3DXO+DEHU'UXJ+HDOWK6HUYLFHV6/+'6RXWK:HVWHUQ6\GQH\/RFDO+HDOWK'LVWULFWAustralia

Dr Janet Lowe
University of Bristol, School of Physiology and Pharmacology United
Kingdomjd.lowe@bristol.ac.uk

Dr Dominique Massotte
IGBMC-CERBM France
massotte@igbmc.fr

Dr Aya Matsui
Vollum Institute / Oregon Health & Science University United States
matsuia@ohsu.edu

0U<RVXNH0DWVXVKLWD
Nagasaki University Japan

paul.haber@sydney.edu.auy-matsushita@nagasaki-u.ac.jp

Ms Asami Hamada
Hoshi University Japan$3URI+LURND]X0L]RJXFKLTohoku Pharmaceutical University Japan

m320@hoshi.ac.jp

mizo@tohoku-pharm.ac.jp

Ms Hendri Hartati
Center for Health Research Universitas Indonesia Indonesia

Ms Sarasa Mohammadi
University of Sydney Australia

emailhendri@yahoo.com

smoh6415@uni.sydney.edu.auMiss Emily Heath
Brain and Mind Research Institute Australia

Mr Md. Ariful Haque Mollik
Prescience Trust Funds United Statesehea7002@uni.sydney.edu.aumollik_bge@live.com

3URI*UDHPH+HQGHUVRQUniversity of Bristol United Kingdom

Ms Shima Momeni
Uppsala University Sweden

graeme.henderson@bris.ac.uk

shima.momeni@farmbio.uu.se

Dr Susan Ingram
Oregon Health & Science University United States

Dr Jose Moron Concepcion
Columbia University United Statesingrams@ohsu.edu

jm3473@columbia.edu

0U<RVKL\XNL,ZDVH
Hoshi University, School of Pharmaceutical Sciences JapanDr Bridin Murnion
Sydney Local Health District Australia

yoshiyuki.i.0905@gmail.com

bmurnion@med.usyd.edu.au

Dr Ernest Jennings
James Cook University Australia

Dr John Muschamp
Harvard Medical School United Statesernest.jennings@jcu.edu.au

jmuschamp@mclean.harvard.edu

Dr Eamonn Kelly
University of Bristol United Kingdom

Mr Karim Nagi
University of Montreal & Sainte-Justine Hospital Research Center
CanadaE.Kelly@bristol.ac.uk

3URI6KLURK.LVKLRND
Wakayama Medical University Japankishioka@wakayama-med.ac.jp

Dr Bronwyn Kivell
Victoria University of Wellington New Zealandbronwyn.kivell@vuw.ac.nz

Ms Alisa Knapman
Australian School of Advanced Medicine Australia
alisa.knapman@students.mq.edu.au

Dr Rakesh Kumar
National Institute of Occupational Health India
rakeshpandey12@gmail.com

Dr Benjamin Lau
Pain Management Research Institute Australia
benjamin.lau@sydney.edu.au

3URI$QGUHZ/DZUHQFH
University of Melbourne Australia$QGUHZ/DZUHQFH#ÁRUH\HGXDX

Ms Erica Levitt
Oregon Health & Science University United States
levitt@ohsu.edu

Ms Minghua Li
Oregon Health & Science University United Stateslimin@ohsu.edu
www.dcconferences.com.au/inrc2013/

karim.nagi@umontreal.ca

Dr Kazuo Nakamoto
Department of Clinical Pharmacy, Kobe Gakuin University, School of
Pharmaceutical Sciences Japankazuo-n@pharm.kobegakuin.ac.jp

3URI,QJULG1\ODQGHU
Uppsala University, Dept Pharmaceutical Biosciences Sweden
Ingrid.Nylander@farmbio.uu.se

Mr Junpei Ohya
Hoshi University, School of Pharmaceutical Sciences Japan
m308@hoshi.ac.jp

Miss Hiroko Ono
Shionogi Co.Ltd. Japan
hiroko_ono@shionogi.co.jp

0V6DUD3DOP
Uppsala University Sweden
sara.palm@farmbio.uu.se'U0DULH2GLOH3DUDW
School of Pharmacy, University of Queensland Australia
m.parat@uq.edu.au'U8OOD3HWDMD5HSR
University of Oulu, Institute of Biomedicine, Department of Anatomy andCell Biology Finland
XOODSHWDMDUHSR#RXOXÀ61

DELEGATE LIST

3URI&KDLP3LFNTel Aviv University Israel

Mr Jacob Thomas
University of Adeladie Australia

chagi.pick@gmail.com

jacob.thomas@adelaide.edu.au'U*UDFLHOD3LQH\UR
Ste-Justine Hospital Canada

Miss Georgina Thompson
Monash Institute of Pharmacological Sciences Australia

JUDFLHODSLQH\URÀOSR#XPRQWUHDOFD

georgina.thompson@monash.edu'U6DEDULQDK3UDVHW\R
Center for Health Research Universitas Indonesia Indonesia

Dr Lawrence Toll
Torrey Pines Institute for Molecular Studies United Statessabarinahprasetyo@gmail.com

ltoll@tpims.org

Mrs Rachel Reid
Washington State University United States

Ms Karen Tonsfeldt
Oregon Health & Science University United Statesrachel.reid@email.wsu.edu

tonsfelk@ohsu.edu

Mrs Louise Rohloff
Mundipharma Australia Australia

Dr Tuan Trang
University of Calgary Canadalouise.rohloff@mundipharma.com.au

trangt@ucalgary.ca

Ms Mahsa Sadeghi
University of Sydney Australia

Dr John Traynor
University of Michigan United States

msad1371@uni.sydney.edu.au

jtraynor@umich.edu

Ms Marina Santiago
Australian School of Advanced Medicine Australia

Dr Thomas Tzschentke
Grunenthal Germany

MARINA.JUNQUEIRA-SANTIAGO@STUDENTS.MQ.EDU.AU

thomas.tzschentke@grunenthal.com

Mr Kazumasu Sasaki
Tohoku University Japan

0U1DRNL8]DZD
Department of Toxicology, Hoshi University School of Pharmacy andPharmaceutical Sciences Japank-sasaki@med.tohoku.ac.jp

3URI6WHIDQ6FKXO]
Institute of Pharmacology, Jena University Hospital Germany
stefan.schulz@mti.uni-jena.de

Dr Marina Shamanskaya
Chita Medical Academy Russian Federation
marshuman@mail.ru

Dr David Sherry
Dept. Cell Biology, OU College of Medicine, OUHSC United States
david-sherry@ouhsc.edu

Ms Setareh Sianati
University of Sydney Australia
ssia7278@uni.sydney.edu.au

3URI0DUHH6PLWKThe University of Queensland Australia
maree.smith@uq.edu.au

3URI$QGUHZ6RPRJ\L
University of Adelaide Australia
andrew.somogyi@adelaide.edu.au

Dr Ichiro Sora
Kobe University Japanichirosora@yahoo.co.jp

Dr Alfreda Stadlin
Chungbuk National University Korea (South)
astadlin@chungbuk.ac.kr

Dr Kelly Standifer
Dept Pharmaceutical Sciences, OU College of Pharmacy, OUHSC UnitedStatesm307@hoshi.ac.jp

3URI5LWD9DOHQWLQRThe Children's Hospital of Philadelphia United States
rjv@mail.med.upenn.edu

Dr Christopher Vaughan
University of Sydney Australia
chris.vaughan@sydney.edu.au

3URI:HQG\:DOZ\Q
University of California, Los Angeles United States
wwalwyn@ucla.edu

0U<RVKLND]X:DWDQDEH
Nippon Chemiphar Japan
yoshikazu-watanabe@chemiphar.co.jp

3URIHVVRU*HRUJH:LOFR[University of Minnesota United Statesgeorge@umn.edu'U$GULDQQH:LOVRQ3RH
University of Sydney Australia
adrianne.poe@sydney.edu.au'U7RPLR<DPDNDZD
Nippon Chemiphar Co.,Ltd. Japant-yamakawa@chemiphar.co.jp

3URI6KX/XQJ<DQJ
Crime Research Center, National Chung Cheng University Taiwan
crmsly@ccu.edu.tw'U$UVDODQ<RXVXIUniversity of Sydney Australia

kelly-standifer@ouhsc.edu

arsalan.yousuf@sydney.edu.au

3URI7VXWRPX6X]XNL
Hoshi University, School of Pharmaceutical Sciences Japan

3URI-DPHV=DGLQD
VA/Tulane University United States

suzuki@hoshi.ac.jp

jzadina@tulane.edu'U3DR/XK7DR
National Health Research Institutes Taiwan

3URI;X=KDQJ
Institute of Neuroscience, Chinese Academy of Sciences China

pltao@nhri.org.tw[X]KDQJ#LRQDFFQ

3URI%UDG7D\ORU
University of Kentucky United States
brad.taylor@uky.edu
62International Narcotics Research Conference 2013

INDEXA
Agus Dwi Setiawan
Alewood, P.F.
Arttamangkul, S.
Atwood, B.K.

H
32, 3833, 42
12
24

B
Bagley, E.
Bailey. C.P.
Baimel, C.
Balleine, B.
Bao, L. 32, 39
Barak, Y.
Beaudry, H.
Bergström, L.
Bhatnagar, S.
Birdsong, W.T.
Borgland, S.L.
Brown, R.M.
Burlingame, A.L.

24
13
28
7
35, 5434, 4735, 52
11, 36, 59
12
28
29
12

C
Canals, M.
Chaijale, N.
Chiang, Huai-Hsiao.
Christie, M.J.
Christopoulos, A.
Comer, S.D.
Connor, M.
Cooper, Z.D.
Cote, T.E.
Cox, B.M.
Curtis A.L.

21, 33, 44
11, 36, 59
32
31
7, 21, 33, 44
17
10, 36, 58
17
25
25
11, 36, 59

D
Daoura, L.
Doll, C.

34, 48
12F
Fujii, H.

45

G
Gendron, L.
Georganta, E.
Georgoussi, Z.
Gerreau, R. IV
Gouty, S.
Granholm, L.

26
32, 4032, 4014
25
34, 48www.dcconferences.com.au/inrc2013/

Hartati, H.
Hayashida, K.
Headrick, J.P.
Henderson, G.
He,Y 34, 46
Hirayama, S.

32, 3745
18
7, 32, 41
45

I
Ikedae, K.
Illing, S.
Ingram, S.L.
Ismail, A.
Itoh, T.
Iwase, Y.
Iwata, N.

36, 5512
15, 32, 40, 41
32, 3736, 5635, 5035, 50J
Jeong, H.J.
Johnson, K.W.
Jones, J.D.
Jupp, B.
Just, S. 12

33, 4517
17
29

K
Kelly, E.
Khoo, S.Y.
Kiguchi, N.
Kim, A.
Kim, J.H.
Kishioka, S.
Knapman, A.
Kobayashi, Y.
Kotowski,S.J.
Kupferschmidt, D.A.

20, 32, 41
29
35, 5129
29
35, 51
10, 36, 58
35, 51
12
24

LLau, B.K.
Lau,E.K.
Lawrence, A.J.
Li, M. 32, 40, 41
Li. M 15
Lovinger, D.M.
Lowe, J.D.

15
12
29
24
13, 32, 41

63

INDEX

M
Machelska, H.
Mann,A.
Manubay, J.
Massotte, D.
Masukawa, D.
Matsushita, Y.
Metz, V.
Miess, E.
Mitchell, V.
Mogali, S.
Mokri, A.
Mollik, A.H. Md.
Momeni, S.
Mori, T.
Mosberg, H.I.
Murata, A.
Muschamp, J.W. Ph.D.

33
12
17
8
36, 5635, 5117
12
33, 4517
1032, 33, 38, 41
35, 5235, 36, 50, 56, 57
33, 4435, 5028

NNadjib, M.
Nagase, H.
Narita, M.
Narita, M.,
Nemoto, T.
Nockemann, D.
Nylander, I.

32, 37
45
14
14
45
33
34, 35, 48, 53O
Ohya, J.

35, 50P
Palm, S.
Parat, M.
Peart, J.N.
Petäjä-Repo, U.
Pineyro, G.
Prasetyo, S.

34, 35, 48, 53
49
18
26
20
32, 37, 38

R
Reid, R.A.
Roman, E.
Rubovitch, V.

64

32, 3815, 32, 40, 41
35, 36, 50, 57
17
35, 36, 50, 56

TTaylor, B.
Thompson, G.
Tokuyama, S.
Tonsfeldt, K.J.
Trang, T.
Trester-Zedlitz,M.
Trinidad, J.C.

1221, 33, 44
36
15, 32, 40, 41
17
12
12

U
Ueda, H.
Uhl, G.R.
Uzawa, N.

35, 51
36
35, 36, 50, 57

V
Valentino, R.J.
Vaughan, C.
Vaughan, C.W.
von Zastrow, M.
Vosburg, S.K.

11, 36, 59
33, 4515
12
17

W
Wakida, N.
Wang, Y.
Wang, Z
Watanabe, C.
Watanabe, M.
Williams, J.T.
Wilson-Poe, A.

35, 5132, 3934, 46
35
34
13
30

X
Xiong, W.

24

Y
35, 5434, 35, 48, 52
35, 54

S
Sadeghi, M.
Saeki, T.
Saika, F.
Saitou, Y.
Schulz, S.
See Hoe, L.E.
Shamanskaya, M.G.
Shibasaki, M.
Silveira, J.T.
Smith, M.T.
Snyder, K.

SubarkahSuchland, K.L.
Sugiyama, K.
Sullivan, M.A.
Suzuki, T.

Yamamoto, C.
Yamashita. A.
Yang, Shu-Lung

35, 5114
11

Z
31
35, 36, 50, 56
35, 5136, 5712
18
32, 3935, 36, 50, 56, 57
25
30
11, 36, 59

Zhang, X.

8International Narcotics Research Conference 2013

NOTES

www.dcconferences.com.au/inrc2013/65

NOTES

66

International Narcotics Research Conference 2013

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