Poster Session Abstracts
The Joint Symposium of WPI-IIIS, Ph.D. Program in Humanics,
and 36th Takamine Conference
Fusion of Biomedical and Physical/Informational Sciences in Neurobiology

Data Blitz by Poster Presenters

15:40 – 16:10

Poster Session

16:10 – 16:50

Poster list
Title

Page

#

Presenter

P01

Seioh Ezaki

P02

Lucas Rebelo
Dal'Bello

Maladaptation in a redundant motor task is caused by history dependency of error-

P03

Taisei Sugiyama

Optimizing learning rate by meta-learning to maximize reward in visuomotor learning

2

P04

Ryota Ishikawa

Sensorimotor Learning from Out-of-Body Viewpoint via the Avatar in Virtual Reality

2

P05

Kazumasa Horie

MC-SleepNet: Large-scale Sleep Stage Scoring in Mice by Deep Neural Networks

3

P06

Liqin Cao

Characterization of sleep architecture in CDKL5 knockout mice

3

P07

Kanako Iwasaki

P08

Fuyuki Asano

Sleep/wakefulness in neuron type-specific SIK3-deficient mice

4

P09

Shinya Nakata

The role of SIK3 kinase activity in sleep/wakefulness

5

P10

Minjeong Park

Sleep/wake behavior of mice lacking PKA phosphorylation site in SIK1 and SIK2

5

P11

Sena Hatori

Functional analysis of SIK3 in Drosophila

6

P12

Yukiko Namekawa

P13

Emi Hasegawa

Roles of dopamine signaling in the amygdala in cataplexy7

P14

Shuntaro Uchida

Delineation of Neural circuits that regulate Muscle Tone during REM sleep

7

P15

Sakura Eri Maezono

Abnormal Sleep and Cognitive ability in an App Knock-in Mouse Model of Alzheimer’s Disease

8

P16

GoEun Han

Slow Wave Sleep and Sleep Need Resolution

8

P17

Sumire Matsumoto

P18

Yoshifumi Arai

P19

Deependra Kumar

The adult-born neuron activity underlying memory consolidation during sleep

10

P20

Pablo Vergara

Ensemble activities of adult-born neurons in memory consolidation during sleep

11

P21

Satomi Okabe

The Effect of Olfactory Stimulation during REM Sleep on Dreams

11

P22

Jaehoon Seol

P23

Tohru M. Takahashi

P24

Shingo Soya

P25

Ai Miyasaka

P26

Kai Yashiro

Neural dynamics of VTA dopaminergic neurons in female sexual behavior

14

P27

Can Liu

Novel neural circuits for innate fear induced defensive responses

14

P28

Akihisa Tokuda

The functional role of delta-opioid system on chronic exposure to social defeat stress

15

P29

Mengxi Yun

Analysis of gait motion change by intervention using robot suit HAL in acute and chronic stage
myelopathy patients decompression surgery
backpropagation capability

Increased non-REM sleep in mice expressing mutant Sik3 in postnatal neurons using newly
developed Synapsin1CreERT2 mice

In vitro and in vivo pharmacology of small-molecule orexin receptor agonists for treatment of
narcolepsy

Neuronal activity of OFF period-like phase during optogenetically evoked cortical response in
SWS
Firing property of the neurons in the medulla involved in sleep/wake cycles and autonomic
nervous system

Effects of types of low intensity physical activity (exercise/housework) in the evening on sleep inolder female adults
Neural Mechanism for Hibernation
Deciphering a novel neural circuit in the central amygdala implicated in regulation of social
behavior
Decoding the sexual behavior of male mice by capturing the dopamine dynamics / Developmentof the automated analyzing system for social behavior

Signal dynamics corresponding to value-to-choice transformation in midbrain dopamine neurons
and orbitofrontal neurons during economic decision-making in monkeys

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6

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15

P01

P02

Analysis of gait motion change byintervention using robot suit HAL in acuteand chronic stage myelopathy patients
decompression surgery

Maladaptation in a redundant motor task iscaused by history dependency of errorbackpropagation capability
Dal’Bello, Lucas R.1, Izawa, Jun2

Seioh Ezaki1, Hideki Kadone2, Shigeki Kubota3,
Yukiko Shimizu3, Tetsuya Abe1, Kousei Miura1,
Yasushi Hada3, Masao Koda1, Masashi Yamazaki1

１

Ph.D. Program in Empowerment Informatics, University ofTsukuba2
Faculty of Engineering, Information and Systems, University ofTsukuba1

Department of Orthopaedic Surgery, Faculty of Medicine,
University of Tsukuba
2
Center for Innovative Medicine and Engineering, University ofTsukuba Hospital3
Department of Rehabilitation Medicine, University of Tsukuba
Hospital

A fundamental, yet poorly understood problem inhuman motor learning – as well as motor recovery – isredundancy: the dimensionality of the corticalrepresentation of motor commands is far larger than thatof the end-effector. However, traditional motor
adaptation studies rely on experimental paradigms that
do not take redundancy into account.
A computational theory called distal-learning predicts
that the brain learns a structure of how to transform avisual error in the task space into an appropriate motor
adaptation signal (called error-backpropagation)
simultaneously with the learning of a controller.
We examine this prediction in a redundant motor task.
We define two perpendicular control environments, A
and B, in a multidimensional motor command space(task-relevant motor commands in A are task-irrelevantin B, and vice-versa), using hand gestures (acquiredwith a 14-DOF data glove) to control a cursor in a 2D
screen. Subjects experience either control environment
A for two days (AA group, N = 4) or A on day one and
B on day two (AB group, N = 4). Learning to manipulate
the cursor progressed well on both conditions and days,
but AB group subjects could not adequately learn theerror-backpropagation structure of B on day two, whichgoes against the prediction of the distal-learning theory.
Our result suggests that forming the errorbackpropagation structure is strongly influenced by theprevious experience on a different control environment,
which may lead to maladaptation in a redundant motor
learning problem such as motor recovery after stroke.

The ossification of the Posterior Longitudinal Ligament(OPLL) is a degenerative spine disease that can cause
motor disorder. Even after appropriate surgicaldecompression, there are some residual motor
impairments could be left. The Hybrid Assistive Limb
(HAL) is a wearable powered suit to assist and support
users voluntary control of hip and knee joint motion by
detecting bioelectric signals from surface of their skinand from force/pressure sensors in the shoes during their
movement. In the current study, the HAL intervention
was applied to 5 acute and 7 chronic phase of OPLL
patients
who
generated
myelopathy
after
decompression surgery, and their gait before and after
intervention was compared.There were significant
improvements in gait speed, cadence, stride length,
swing time. Peak comparison of joint angles of leg wasused to evaluate the limb movement during the gait;
Range of movement (ROM) of hip joints, knee joints,
foot joints, and toe lift have increased significantly, anddouble knee action has appeared after HAL intervention.
ROM of Elevation angle in thigh, shank, and foot has
enlarged increased significantly. Improvement in all theangles above except ROM of foot joint was observed forthe chronic patients. HAL training enable patients to
do repetitive training without giving stress on knee
joints, so that lead them to learn and control knee
muscles correctly. Importantly, instead of gaining knee
hypertension that is common gait impairment for OPLL,
double knee action has appeared after the HAL
intervention and it leads to smoother and healthier gait
motion.

1

P03

P04

Optimizing learning rate by meta-learning tomaximize reward in visuomotor learning

Sensorimotor Learning from Out-of-Body
Viewpoint via the Avatar in Virtual Reality

Taisei Sugiyama1, Nicolas Schweighofer2, Jun Izawa3

Ryota Ishikawa*, Kyohei Sasaki, Saho Ayabe, and Jun
Izawa

１

Empowerment Informatics, University of Tsukuba
２
Biokinesiology and Physical Therapy, University of Southern
California
３
Engineering, Information and Systems, University of Tsukuba*

Master program in Comprehensive Human Sciences, Universityof Tsukuba

It is effective to use mental imagery for or see one’s own
body movement in order to improve motor skills. This
perspective is stated as an out-of-body viewpoint.
Recently engineers implemented new devices by which
it is possible to obtain visual feedback of the user’s
posture and body movement (i.e. “self-image”) via theavatar in a virtual reality (VR) environment [Hamanishi
et al., 2019]. This system has an advantage inmonitoring the self-image on using mirrors or video
recording since it can provide online visual feedback
from free viewpoint. However, it remains unclear how
and to what extent the out-of-body viewpoint
contributes to sensorimotor learning. Here we devised anovel visuomotor adaptation task in the VR space where
the participants observed a self-body motion from theout-of-body viewpoint (i.e. the third-person
perspective) via the avatar. In addition, we asked theparticipants to conduct another cognitive task
measuring the individual’s ability to take other’s pointof view (Spatial Perspective Taking, SPT). Estimating
parameters involved in the state-space model for
sensorimotor learning based on the dual-rate learning
process (Smith et al., 2006), we examined the relationsof each parameter and the performance of the cognitivetask. The results indicated that in the computationalmodel SPT was important for processing errors, but notfor learning or retention factors.

Both human and animals accelerate learning in a varietyof tasks and environments, yet its underlying
mechanism is unclear. In machine learning, regulationof learning rate is achieved by reinforcement metalearning to maximize reward. If the same algorithm
underlies regulation of motor learning rate, therelationship between motor learning and reward should
determine how a learner changes learning rate. Here, we
implemented a reinforcement meta-learning problem ina visuomotor arm-shooting task. Specifically, humanparticipants first observed sensory prediction error
induced by rotation of cursor feedback during
movement in one trial, and then the degree of learningfrom the error (aftereffect) was evaluated with monetary
feedback in subsequent trials. By manipulating therelationship between learning and reward, we
demonstrated that learning was accelerated overtraining when fast learning yielded more reward
compared to when fast learning yielded less reward (p= .01), and the effect remained transiently after reward
was removed (p = .04). The results support regulation of
motor learning rate by reinforcement learning,suggesting that people optimize learning rate bylearning (meta-learning) to maximize reward. As
growing evidence from neuroimaging research suggeststhat the cerebellum is sensitive to not only sensory error
for motor learning but also reward, we speculate that the
cerebellum may interact with the reinforcement learning
system (e.g., cortico-basal ganglia) for tuning motor
learning rate and increasing reward from theenvironment.

2

P05

P06

MC-SleepNet: Large-scale Sleep Stage
Scoring in Mice by Deep Neural Networks

Characterization of sleep architecture in
CDKL5 knockout mice

Kazumasa Horie, Hiroyuki Kitagawa

Liqin Cao1, Jing Ma1, Tingting Lou1, Zhiqang Wang1,
Qinghua Liu1, 2, 3Center for Computational Sciences, University of Tsukuba

1

International Institute of Integrative Sleep Medicine (WPIIIIS), University of Tsukuba, Japan
2
University of Texas Southwestern Medical Center, USA
3
National Institute of Biological Sciences, China

Automated sleep stage scoring for mice is in highdemand for sleep research, since manual scoring
requires considerable human expertise and efforts. The
existing automated scoring methods do not provide thescoring accuracy required for practical use. In addition,
the performance of such methods has generally been
evaluated using rather small-scale datasets, and theirrobustness against individual differences and noise has
not been adequately verified. This research proposes anovel automated scoring method named “MCSleepNet”, which combines two types of deep neuralnetworks. Then, we evaluate its performance using alarge-scale dataset that contains 4,200 biological signal
records of mice. The experimental results show that
MC-SleepNet can automatically score sleep stages withan accuracy of 96.7% and kappa statistic of 0.94. In
addition, we confirm that the scoring accuracy does not
significantly decrease even if the target biologicalsignals are noisy. These results suggest that MCSleepNet is very robust against individual differences
and noise. To the best of our knowledge, evaluations
using such a large-scale dataset (containing 4,200
records) and high scoring accuracy (96.7%) have notbeen reported in previous related studies.

CDKL5 deficiency disorder (CDD) is a devastating Xlinked neurodevelopmental disorder caused bypathogenic mutations in the cyclin-dependent kinaselike 5 (CDKL5) gene. Patients with CDD display avariety of clinic symptoms that include early-onset
refractory seizures in the first months of life,
developmental delay, intellectual and motor disabilities,
cortical visual impairment, and autistic-like symptoms.
The majority (>86%) of patients have severe sleep
problems, which greatly affect the quality of life forpatients and their families. To gain insight into thepathogenesis of CDKL5-related diseases, several
CDKL5-deficient mouse models have been generated.
These mice recapitulated many aspects of human CDD
symptoms, and exhibited impaired learning and memory,
motor and visual dysfunction, and autistic-likephenotypes. However, little is known about their sleep
phenotypes. We characterized baseline sleep/wakepatterns and recovery sleep following sleep deprivationin adult Cdkl5 knockout male mice and their wild-type(WT) littermates by electroencephalography andelectromyography. CDKL5-null mice exhibited
increased sleep latency, less overall sleep time, shorter
sleep episode duration and frequent awakenings
compared to WT mice which resemble sleep
disturbances observed in human CDD patients. Our
results suggest that the Cdkl5 knockout mouse model
may be a useful genetic model for studying sleep
disruptions in CDD patients.

3

P07

P08

Increased non-REM sleep in mice expressing
mutant Sik3 in postnatal neurons using
newly developed Synapsin1CreERT2 mice

Sleep/wakefulness in neuron type-specific
SIK3-deficient mice

1

Fuyuki Asano1, Tomoyuki Fujiyama1, Chika Miyoshi1,
Noriko Hotta-Hirashima1, Miyo Kakizaki1, Aya Ikkyu1,
Satomi Kanno1, Shinya Nakata1, Nodoka Asama1, Seiya
Mizuno2, Fumihiro Sugiyama2, Satoru Takahashi1,2,
Hiromasa Funato1,3, Masashi Yanagisawa1

1

Kanako Iwasaki , Tomoyuki Fujiyama , Shinya
Nakata1, Park Minjeong1, Noriko Hotta-Hirashima,
Fumihiro Sugiyama2, Seiya Mizuno2, Satoru
Takahashi2, Hiromasa Funato1,3, Masashi
Yanagisawa1,4,5

１

WPI-IIIS, University of Tsukuba, Ibaraki, Japan
Faculty of Medicine, University of Tsukuba, Ibaraki, Japan３
Dep Anat, Toho University, Tokyo, Japan
２

1

International Institute for Integrative Sleep Medicine (WPIIIIS), University of Tsukuba, Tsukuba, Ibaraki, Japan
2
Laboratory Animal Resource Center, University of Tsukuba,
Tsukuba, Ibaraki, Japan3
Department of Anatomy, Faculty of Medicine, Toho University,
Ota-ku, Tokyo, Japan
4
Department of Molecular Genetics, University of Texas
Southwestern Medical Center, Dallas, TX, USA
5Life Science Center, Tsukuba Advanced Research Alliance,
University of Tsukuba, Tsukuba, Ibaraki, Japan

Sleep is a fundamental behavior conserved fromvertebrates to invertebrates. Although the two-processmodel has been proposed as a conceptual framework ofsleep regulation, the molecular mechanism of
sleep/wake regulation and neural substrate of sleep needremain unknown. SIK3 is a member of AMP-activatedprotein kinase (AMPK) family and is associated withsleep need. Gain-of-function Sik3 gene mutant miceincrease sleep need and time spent in NREM sleep.
Almost all systemic SIK3-deficient mice die neonatallyand very few survived mice suffered from several
growth retardation and malnutrition, Thus, these mice
are not suitable for the sleep/wake analysis.
Here, we examined whether neuronal SIK3-deficiency
affects sleep/wakefulness and which neuronal groups
are responsible for sleep/wake regulation using newly
generated Sik3-flox mice. Pan-neuronal SIK3-deficient
mice were healthy. We examined sleep/wakefulness
through EEG/EMG analysis. We also examinedcircadian behavior using a running-wheel under theconstant darkness. SIK3-deficiency in glutamatergic
neurons resulted in decreased NREMS delta power andtime spent in NREMS. SIK3-deficiency in GABAergic
neurons prolonged the free-running period while delta
power in NREMS was not affected. These results
indicate that the neuronal group responsible for sleep
regulation via SIK3 is different from those for circadianbehavior.

Sleep amount is regulated in a homeostatic manner sothat the drive for sleep become stronger after sleep
deprivation. Despite empirical evidence of homeostatic
regulation of sleep, the molecular and neuralmechanism determine sleep need or sleep amountremains unknown. Recently, we identified a gain-offunction Sleepy mutation in salt-inducible kinase 3(SIK3) that increases the sleep need and time spent in
non-REM sleep. The Sleepy mutant mice investigated inprevious studies systemically expressed the mutant
allele of Sik3 in the germline. Therefore, it remained tobe elucidated whether neurons or other types of cells are
responsible for the increased non-REM sleep.
Additionally, we could not deny the possibility thatmutant Sik3 expression in the fetal brain decides thetime spent in non-REM sleep in adult mice.
Here, we investigated sleep/wake behavior in miceexpressing Sleepy mutant Sik3 in postnatal neurons. We
newly generated Synapsin1CreERT2 mice to manipulate atarget gene in the neurons after tamoxifen
administration.
HistologicalexaminationofSynapsin1CreERT2 mice showed tamoxifen induced
recombination in neurons and a few peripheral tissues.
Synapsin1CreERT2 mice were then crossed with Sik3flox/+
mice, which express Sleepy mutant Sik3 in a Credependent manner. EEG/EMG-based sleep staging
of Synapsin1CreERT2; SIK3flox/+ mice revealed thattamoxifen-treated Synapsin1CreERT2; SIK3flox/+ mice
exhibited increased non-REM sleep compared tocontrol mice. Our results demonstrated that the mutant
SIK3 regulates sleep amount by acting in postnatal
neurons.
4

P09

P10

The role of SIK3 kinase activity insleep/wakefulness

Sleep/wake behavior of mice lacking PKA
phosphorylation site in SIK1 and SIK2

Shinya Nakata1,2, Tomoyuki Fujiyama2, Chika Miyoshi2,
Aya Ikkyu2, Fuyuki Asano2, Haruna Komiya2, Noriko
Hotta-Hirashima2, Seiya Mizuno3, Fumihiro Sugiyama3,
Satoru Takahashi3, Hiromasa Funato2,4, Masashi
Yanagisawa2

Minjeong Park1, Chika Miyoshi1, Tomoyuki Fujiyama1,
Miyo Kakizaki1, Aya Ikkyu1, Takato Honda1, Jinhwan
Choi1, Fuyuki Asano1, Seiya Mizuno 2, Satoru
Takahashi 2, Hiromasa Funato1,3, Masashi Yanagisawa1,4
1International Institute for Integrative Sleep Medicine (WPI-IIIS),
University of Tsukuba
2
Laboratory Animal Resource Center, University of Tsukuba
3
Department of Anatomy, Toho University School of Medicine
4
University of Texas, Southwestern Medical Center

1

Ph.D program in Humanics, University of Tsukuba
2
WPI-IIIS, University of Tsukuba
3
Faculty of Medicine, University of Tsukuba
4
Department of Anatomy, Toho University

The mouse pedigree, Sleepy, which has a splicing
mutation in Sik3 resulting in the skipping of exon 13,
shows prolonged non-REM sleep time and increased
sleep need. Since our results showed a hypomorphic
Sik3 mutation in mice and Drosophila resulted in adecrease in sleep time, the Sleepy mutation seems to bea gain-of-function allele. Moreover, it has been reported
that SIK3 kinase activity is tightly linked withphosphorylation of Thr221 (T221) in the kinase domain
T-loop of SIK3. Interestingly, the phosphorylation of
T221 was increased in wild-type mice after sleep
deprivation, suggesting that SIK3 kinase activity
increases in mice which have a higher sleep need.
However, whether the SIK3 kinase activity is involved
in sleep regulation is still unknown.
Here, we investigated the role of SIK3 kinase activity inits phosphorylation status and established geneticallymodified Sik3T221E mice whose SIK3 is constitutively
active. First, we examined the phosphorylation status
and physical interaction of mutant SIK3 proteins using
HEK293T cells. For further investigation, we havemade Sik3T221E mice using CRISPR/Cas9 system. We
confirmed the establishment of Sik3T221E mutant mice by
dCAPS genotyping and direct sequencing.

We previously identified a kinase, SIK3, as an important
sleep regulator through screening of randomly
mutagenized mice. Mice that express mutant SIK3
lacking the 52 amino acids encoded by exon 13 showeda decrease in wake time and an increase in NREM sleep
time. SIK3 is an AMPK-family protein kinase
containing a well-conserved protein kinase A (PKA)phosphorylation site, serine 551. The skipping of exon
13 results in a deletion of 52 amino acids including S551.
Also, Sik3 S551A knock-in mice showed reduced total
wake time and increased sleep need. These results
suggest that PKA-SIK3 pathway is involved in theregulation of sleep/wake behavior.
The SIK family is composed of SIK1, SIK2, and SIK3.
Importantly, an S551-equivalent serine residue isconserved at S577 in SIK1 and at S587 in SIK2. To
examine whether the phosphorylation of S577 of SIK1
and S587 of SIK2 is required for proper sleep/wakebehavior, we generated mutant mice in which SIK1
S577 and SIK2 S587 were substituted by alanine
through the CRISPR/Cas9 method. Both Sik1 S577A
mice and Sik2 S587A mice showed increased NREM
sleep delta power, an index for sleep need to a smaller
extent than Sik3 S551A mice. It can be explained by thelower expression of Sik1 and Sik2 than Sik3 in the brain.
Furthermore, Sik1 S577A mice showed reduced waketime but showed normal circadian behavior and reentrainment to a new circadian rhythm. These findings
indicate the PKA recognition sites of SIK family are
required for regulation of sleep need.

5

P11

P12

Functional analysis of SIK3 in Drosophila

In vitro and in vivo pharmacology of smallmolecule orexin receptor agonists fortreatment of narcolepsy.

Sena Hatori1, Riho Kobayashi1, Hiroyuki Nakagawa1,
Jun Tomita1, Hiromasa Funato2, Masashi Yanagisawa2,
Kazuhiko Kume1

Yukiko Namekawa*, Yoko Irukayama-Tomobe,
Yukiko Ishikawa, Yamaguchi Takuto, Yasuhiro Ogawa,
Tsuyoshi Nemoto, Hiromu Tanaka, Hikari Yamamoto,
Tito Akindele, Tsuyoshi Saitoh, Hiroshi Nagase andMasashi Yanagisawa

1

Graduate School of Pharmaceutical Sciences, Dept.
Neuropharmacology, Nagoya City University
2
International Institute for Integrative Sleep Medicine (WPI-IIIS),
University of Tsukuba

The amount of sleep is regulated by homeostasis, but theunderlying molecular mechanism is still unclear.
However, recent study identified Sleepy1 mutation.
Sleepy1 mutation was discovered in an unbiased genetic
screening of more than 8000 mice and was identified as
an exon-skip mutation of Sik3, salt inducible kinasegene. This mutation dramatically increased total sleep
time in mice (Funato et al. Nature, 2016). Sik3 has aserine residue in the skipped exon, which is
phosphorylated by PKA and conserved in Drosophila.
In this study, we used phosphorylation-defective SIK3,
in which the serine residue is substituted to an alanine
residue (SIK3SA), in order to examine the function of
SIK3 phosphorylation in sleep. Using GAL4-UAS
system and GeneSwitch system, we found pan-neuronalover-expression of SIK3SA and conditional overexpression of SIK3SA increased sleep. The effects ofthis mutant gene were more remarkable in constant dark(DD) conditions, especially in subjective daytime, thanin light-dark (LD) conditions. In order to localize where
SIK3SA functions to regulate sleep, we expressed
SIK3SA only in the subsets of neurons using various
GAL4 drivers. Its over-expression in PDF neurons,
which are master clock neurons also resulted in increasein sleep. These data indicate SIK3SA modulates sleep
amount controlling the functions of PDF neurons. Next,
we investigated SIK3 target genes and identified a novel
sleep regulating gene. Overexpression of this gene in all
neurons increased sleep. Now, we analyzed the detailed
functions of this gene and interactions with Sik3.

*

Yanagisawa/Funato laboratory, International Institute forIntegrative Sleep Medicine (WPI-IIIS), University of Tsukuba

Sleep/wakefulness is regulated by orexin, aneuropeptide produced by neurons exclusively
localized in the lateral hypothalamus. Orexin deficiency
causes narcolepsy-cataplexy characterized by excessive
sleepiness, sleep/wake fragmentation and cataplexy.
Orexin acts on two receptors, OX1R and OX2R, andOX2R is the main receptor regulating sleep; the role of
OX1R is less clear. As orexin cannot pass the bloodbrain barrier, the peptide is difficult to use as a clinicaldrug. Therefore, small-molecule orexin receptor
agonists, especially OX2R agonists, are expected to be anovel therapy for narcolepsy-cataplexy. We previously
showed that YNT-185, an OX2R-selective agonist,
ameliorates narcolepsy-cataplexy symptoms in mousemodels when peripherally (i.p.) administered. However,
the effective dose of this compound for oral
administration (p.o.) was too high.
Here we further optimized YNT-185 (EC50 for OX2R
≈ 28 nM) and produced 2 different types of agonists; a
OX2R-selective agonist, YNT-X (EC50 for OX2R ≈ 1.1
nM by Ca assay) and a OX1R/OX2R agonist, YNT-Y2(EC50 for OX1R ≈ 3.7 nM, EC50 for OX2R ≈ 0.9 nM).
Oral administration of these compounds increased waketime in wild-type mice in a dose-dependent manner.
Their effective doses are several hundred times lower
than YNT-185. These effects were not detected in orexin
receptor-deficient mice. In addition, PO administrationof
YNT-Y2ameliorated
narcolepsy-cataplexy
symptoms in orexin-deficient mice. Previous reportssuggest the possibility that OX1R signal may be
important for consolidation of wakefulness. We will beable to dissect the role of orexin receptor subtypes innarcolepsy treatment by using these two types of orexin
agonists.

6

P13

P14

Roles of dopamine signaling in the amygdalain cataplexy

Delineation of Neural circuits that regulate
Muscle Tone during REM sleep

Emi Hasegawa1, Ai Miyasaka1, Katsuyasu Sakurai1,
Takeshi Sakurai1,2,3

Shuntaro Uchida, Shingo Soya, Takeshi Sakurai
International Institute for Integrative Sleep Medicine (WPI-IIIS),
University of Tsukuba

1

International Institute for Integrative Sleep Medicine (WPIIIIS)
2
Faculty of Medicine
3Life Science Center for Tsukuba Advanced Research Alliance(TARA), University of Tsukuba, JapanOne of the hallmarks of REM sleep is bilateral muscle
atonia. Cataplexy, a cardinal symptom of narcolepsy, isa sudden weakening of muscle tone triggered by strong
emotions and considered to be a REM-related syndrome.
However, the precise neuronal circuits that regulate
REM atonia and cataplexy are unknown. The purposeof this study is to delineate neuronal circuits that evoke
the REM atonia and examine whether this pathway isalso involved in cataplexy or not. Recent studies inrodents showed that inhibitory neurons in theventromedial medulla (VMM) are responsible forgenerating the muscle atonia during REM sleep.
Consistently, we observed obvious increases in muscle
tone and body movements during REM sleep in mice
when we inhibited glycinergic neurons in the VMM(GlyVMM neurons) by expression of tetanus toxin lightchain. Furthermore, we also found that silencing
GlyVMM neurons significantly decreased both theduration and number of cataplexies in narcolepsy model
mice. Rabies virus mediated retrograde tracing revealed
that neurons located in the brainstem such as thesublaterodorsal tegmental nucleus (SLD) and the
pontine reticular nucleus (Pn) make direct synaptic
contacts with GlyVMM neurons that send projection to
the anterior horn of the spinal cord. We found thatspecific silencing of the glutamatergic neurons in the
SLD (GlutSLD) that project to the VMM resulted in REM
sleep without atonia and decreased time spent in
cataplexy in narcoleptic mice. These findings suggestthat the GlutSLD →GlyVMM pathways are common in
REM atonia and cataplexy.

Although the lack of orexin signaling causes the sleep
disorder narcolepsy, the precise neural mechanisms bywhich orexin neurons prevent narcolepsy remain
unclear. In a previous study, we found that targeted
restoration of orexin receptor expression in the dorsal
raphe nucleus (DR) and in the locus coeruleus (LC) of
mice lacking both of orexin receptors inhibited
cataplexy and pathological fragmentation of
wakefulness (i.e., sleepiness), respectively. These
results suggested that DR serotonergic (DR5HT) and LC
noradrenergic (LCNA) neurons play differential roles in
orexinneuron-dependent
regulationofsleep/wakefulness.
As a next step, we used
optogenetic and chemogenetic approaches todemonstrate that DR5HT neurons suppress cataplexy by
reducing the activity of the basolateral/lateral amygdalathat plays an important role in emotional processing, asconsistent with the fact that strong emotion often
triggers cataplexy. Our results suggest that the orexin
neuron→DR5HT→amygdala pathway is a critical circuit
for preventing cataplexy. Furthermore, we identified aneuronal pathway that induces cataplexy when activatedby optogenetic manipulation. We focused on the reward
system dopamine from the fact that strong emotions,especially positive emotions, often cause cataplexy. The
ventral tegmental area to amygdala pathway is a criticalcircuit for inducing cataplexy. We will discuss the roleof this pathway in emotional processing as well as in
REM-related muscle atonia.

7

P15

P16

Abnormal Sleep and Cognitive ability in an
App Knock-in Mouse Model of Alzheimer’s
DiseaseSlow Wave Sleep and Sleep Need Resolution

Sakura Eri Maezono1,2, Mika Kanuka1, Chika
Tatsuzawa1, Miho Morita1, Mitsuaki Kashiwagi1,
Pimpimon Nondhalee1, Masanori Sakaguchi1, Takashi
Saito3,4, Takaomi Saido3, Yu Hayashi1

1

GoEun Han1, Javier Diaz2, Robert Greene2,3 1 and
Kaspar Vogt2
Ph.D. Program in Human Biology, University of Tsukuba
International Institute for Integrative Sleep Medicine (WPIIIIS), University of Tsukuba
3
Department of Psychiatry, University of Texas Southwestern
Medical Center, Dallas, Texas 75390, USA
2

1

International Institute for Integrative Sleep Medicine (WPIIIIS), University of Tsukuba, Japan
2
School of Integrative and Global Majors, University ofTsukuba, Japan
3
Laboratory for Proteolytic Neuroscience, RIKEN Center for
Brain Science, Japan
4
Department of Neuroscience and Pathobiology, ResearchInstitute of Environmental Medicine, Nagoya University, Japan

Sleep is necessary for survival, however, its
mechanisms are still not fully understood. In sleep, thebrain generates slow electrical signals (1-4 Hz) that canbe recorded in the electro-encephalogram (EEG); thisslow wave activity (SWA) homeostatically increases
with the duration of the waking time before sleep anddecreases during sleep.
SWA is synchronous activity of cortical neurons duringslow wave sleep (SWS), oscillating between silent OFF
and active ON periods. During SWS ON periods,
activity-dependent influx of calcium ions into neurons
through voltage-gated calcium channels (VGCCs) isobserved, but the function of this calcium influx is
unclear.
We hypothesize that neuronal calcium influx is involved
in resolving sleep need, visible as decreases in SWA
during sleep. We used nifedipine, a common
antihypertensive drug that readily penetrates the bloodbrain barrier to block L-type VGCCs, responsible for asignificant fraction of activity-dependent postsynaptic
calcium influx in cortical neurons.
Nifedipine reduced EEG SWA for 1-2 h after
subcutaneous injection in mice at the beginning of thelight phase, when sleep need and SWA are highest. Thisdecrease in SWA was followed by a rebound after the
nifedipine-effect had worn off. Based on these results,
we hypothesize that neuronal calcium influx contributes
to sleep need resolution during SWA and that asignificant fraction is contributed by L-type VGCCs.

Alzheimer’s disease (AD) is the most common
neurodegenerative disease. It is a slow progressing
disease characterized by aggregation of amyloid-beta(Aβ) peptides. In addition to cognitive decline, AD
patients often suffer from poor sleep. These symptoms
not only affect their QOL but also those of the primary
caregivers. Increasing studies support that sleep deficits
can contribute to the progression of AD, consistent withthe roles of sleep in memory and brain homeostasis.
Thus, it is crucial to understand the underlying
mechanisms of sleep disorders in AD.
Here, we unveil the sleep abnormalities exhibited by an
AD mouse model. We chose the App knock-in mouse(Saito et al., Nature Neuroscience, 2014). In this AD
mouse model, Aβ accumulates in a manner similar tohuman AD via mutation in the gene encoding amyloidbeta precursor protein (APP), and thus, various aspectsof human AD are faithfully recapitulated. In these mice,at 6 months of age, when cognitive deficits become
apparent, impairment in various aspects of the sleep
architecture were detected. These phenotypes became
more severe at 12 months of age. Consistent with thenotion that sleep is important for memory consolidation,
the AD mouse model also exhibited learning deficits.
We expect that our results provide a starting point foraddressing how sleep is impaired in AD and how it canaffect cognitive functions.

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Neuronal activity of OFF period-like phaseduring optogenetically evoked corticalresponse in SWS

Firing property of the neurons in the medulla
involved in sleep/wake cycles and autonomic
nervous system

Sumire Matsumoto1,2, Kaoru Ohyama 2,3, Javier Díaz 2,
Kaspar Vogt 2

Y. Arai*, H. Iwata, T. Suzuki, K. Mochizuki, Y. Koyama*

Faculty of Symbiotic Systems Science, Fukushima University

1

Ph.D. Program in Human Biology, School of Integrative andGlobal Majors, University of Tsukuba, Tsukuba, Japan
2
International Institute of Integrative Sleep Medicine (WPI-IIIS),
University of Tsukuba, Tsukuba, Japan
3
Japan Society for the Promotion of Science (JSPS) Research
Fellow
4
Department of Psychiatry & Neuroscience, Peter O’Donnell
Brain Institute, UT Southwestern Medical Center, Dallas, TX,
USA

The medulla is well known for the center of the
cardiovascular regulation. Besides that,the medulla plays crucial roles in the promotion and themaintenance of rapid eye movement (REM) sleep.
During REM sleep, blood pressure (BP) surges andrapid heart rate (HR) increases are observed due to theturbulence of the autonomic nervous system. However,it remains to be known how the neurons in the medulla
regulate sleep/wake cycles and BP fluctuation during
REM sleep.
We recorded single neuronal activity and BP in headrestrained and unanaesthetised rats during sleep/wakecycles. Of 115 neurons recorded, 75.7% (87/115)
showed increased activity during REM sleep, REM
sleep and waking, or Slow-wave sleep and REM sleep,
which we term PS active neurons.
32.2 % of PS active neurons (28/87) started increasing
their activity before the REM sleep onset. They aremainly located from central to the caudal medulla. The
neurons in the caudal parts, which preceded the REM
sleep onset, mostly showed phasic firing, whereas theneurons in the central parts showed tonic firing.
Of 87 PS active neurons, 32.2 % (28) showed firing inclose relation with BP fluctuation during REM sleep,and 75 % of them (21/28) increased their firing rate
before the BP fluctuation. These results suggest thatneurons in the medulla are involved in the regulation of
rapid BP increases during REM sleep.

Slow wave activity (SWA) during slow wave sleep(SWS), recorded from electroencephalograms (EEG) or
local field potential (LFP) is a key indicator of sleep
homeostasis and therefore it likely has a central role insleep. It has been reported that SWA is not only
spontaneously occurred but can also be induced bytranscranial current stimulation (Marshall et al., Nature,2006), magnetic stimulation (Massimini et al., PNAS,
2007), auditory stimulation (Tononi et al., Medicamundi,2010), or electrical stimulation directly delivered tocortex (Vyazovskiy et al., J Neurophysiol. 2009). Here
we investigated the cortical activity induced by brief
optogenetic stimulation during SWS in mice andcompared to the spontaneous OFF (silent) period ofslow wave. The optogenetic stimulation triggered abiphasic response, a brief excitation followed by longsilence of unit activity which was similar to the OFF
period of SWA. However, the cortical neurons showed
less excitability during the stronger evoked-silence
period while larger excitation was observed during
spontaneous OFF period compared to ON period. We
further analyzed the neuronal activity of corticalneurons during the evoked response and found that thelate silent period was most likely to be induced by the
feedback inhibition which is triggered by the preceded
brief excitation.

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DG. After TAM injection, we targeted different ages(~2, 4, and 10-week old) of the ABNs for silencing
during each sleep stage.

The adult-born neuron activity underlying
memory consolidation during sleep
Deependra Kumar1, Iyo Koyanagi1†, Alvaro CarrierRuiz2,3†, Pablo Vergara1†, Sakthivel Srinivasan1†, Yuki
Sugaya2,3, Masatoshi Kasuya1, Tzong-Shiue Yu4,
Kaspar E. Vogt1, Masafumi Muratani5, Takaaki
Ohnishi6, Sima Singh1, Catia M. Teixeira7, Yoan
Chérasse1, Toshie Naoi1, Szu-Han Wang8, Pimpimon
Nondhalee1, Boran A.H. Osman1, Nahoko Kaneko9,
Kazunobu Sawamoto9,10, Steven G. Kernie4, Takeshi
Sakurai1, Thomas J. McHugh11, Masanobu Kano2,3,Masashi Yanagisawa1, Masanori Sakaguchi1

Result: Light-induced silencing of the 4-week old ABN
activity during sleep within the memory consolidation
window impaired memory. It also led to elongation of
ABN dendritic spine neck. Furthermore, no memory
impairment was observed when ~2- or 10-week-old
ABN activity was silenced in the same manner.
Conclusion: We provide causal evidence that theactivity of the young ABNs is necessary for memory
consolidation during sleep.

1

International Institute for Integrative Sleep Medicine (WPI-IIIS),
University of Tsukuba, Tsukuba, Ibaraki, Japan2
Department of Neurophysiology, Graduate School of Medicine,
The University of Tokyo, Tokyo, Japan
3
International Research Center for Neurointelligence (WPIIRCN), The University of Tokyo Institutes for Advanced Study(UTIAS), Tokyo, Japan
4
Department of Pediatrics, Columbia University College ofPhysicians and Surgeons, New York, NY, USA.
5
Department of Genome Biology, Faculty of Medicine, Universityof Tsukuba, Tsukuba, Ibaraki, Japan
6
Graduate School of Information Science and Technology, The
University of Tokyo, Tokyo, Japan
7
Emotional Brain Institute, Nathan Kline Institute, Orangeburg,
NY, USA
8
Centre for Clinical Brain Sciences, University of Edinburgh, UK.
9
Department of Developmental and Regenerative Biology,
Nagoya City University Graduate School of Medical Sciences,
Nagoya, Aichi, Japan
10
Division of Neural Development and Regeneration, National
Institute for Physiological Sciences, Okazaki, Aichi, Japan
11
RIKEN Center for Brain Science, Wako, Saitama, Japan†These authors contributed equally to this work.

Background: The hippocampus plays critical roles inmemory consolidation during sleep, yet its responsible
neural population is still unclear. In the hippocampus,
the dentate gyrus (DG) continuously generate newneurons throughout life. These adult-born neurons(ABNs) are crucial for learning and memory retrieval,but their functions in sleep are completely unknown.
Method: We used the pNestin-CreERT2 driver mice tolabel and manipulate ABNs. Injection of an estrogen
ligand, tamoxifen (TAM), to the mice induced
recombination in the neural progenitor cells to express
Halorhodopsin, a light-sensitive neuronal silencer, in
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Ensemble activities of adult-born neurons inmemory consolidation during sleep

The Effect of Olfactory Stimulation during
REM Sleep on Dreams

Pablo Vergara1, Alvaro Carrier-Ruiz2,3, Deependra
Kumar1, Iyo Koyanagi1, Sakthivel Srinivasan1, Yuki
Sugaya2,3, Yoan Chérasse1, Sima Singh1, Toshie Naoi1,
Takeshi Sakurai1, Masashi Yanagisawa1, Masanobu
Kano2,3, Masanori Sakaguchi1

Satomi Okabe1, Takashi Abe1, Mitsuo Hayashi2, Kazuo
Yamada3,
Hiroko-Mochizuki-Kawai4,
Kazuhiko
5
Fukuda1

International Institute for Integrative Sleep Medicine (WPIIIIS), University of Tsukuba
2Graduate School of Integrated Arts and Sciences, Hiroshima
University
3
Faculty of Human Sciences, University of Tsukuba
4
National Agriculture and Food Research Organization
5
College of Sociology, Edogawa University

1

International Institute for Integrative Sleep Medicine (WPI-IIIS),
University of Tsukuba, Tsukuba, Ibaraki, Japan2
Department of Neurophysiology, Graduate School of Medicine,
The University of Tokyo, Tokyo, Japan
3
International Research Center for Neurointelligence (WPIIRCN), The University of Tokyo Institutes for Advanced Study(UTIAS), Tokyo, Japan

Dreams have been investigated, not only to clarify
clinical problems such as possible associations between
nightmares and depression, but also to understandfundamental processes such as mechanisms of memoryor the purpose of rapid eye movement (REM) sleep.
Compared with other sensory stimulations, olfactorystimuli may be able to be processed in the brain during
dreaming, and without causing arousals. However,
although olfactory perception shows large individual
differences, these individual differences have not been
considered. We therefore conducted two experiments toclarify if individual differences in odor perception affectdream emotionality after olfactory stimulation during
REM sleep. Our investigation of the preference factor
comprised Experiment 1. Study participants weredivided into two groups according to each participant’s
individual preference for Phenylethyl alcohol (rose-likefragrance) odor, i.e. prefer or not-prefer groups. Eachgroup was exposed to control and odor conditions. The
goal of Experiment 2 was to investigate the effects offamiliarity. There were two groups, i.e. familiar andunfamiliar groups. We observed effects of odor on
dreams only in the prefer group (Experiment 1) andfamiliar group (Experiment 2). In addition, for bothgroups, dreams which occurred in odor conditions weremore emotionally negative than those in control
conditions.

The hippocampal dentate gyrus (DG) is one of the few
brain regions that continue to produce new neurons
throughout life. We showed that the adult-born neuron(ABN) activity during sleep is necessary for memory
consolidation (Kumar et al., in revision). Here, we
employ calcium imaging of ABNs under the free
moving condition to reveal the mechanisms of memory
consolidation during sleep. Under home cage conditions,
ABNs display repetitive patterns of neuronal ensemble
activities across sleep/wake periods. Interestingly, theensemble activities are more frequent when the DG
local field potential shows a synchronous oscillatory
activity, theta oscillations. Theta oscillations play
critical roles in the temporal coordination of neuronal
activities (Buzsáki, Neuron 33: 325-40, 2002). We also
examine the ensemble activities during encoding,consolidation, and retrieval periods of the contextual
fear memory. We intend to show our most recent data at
the presentation.

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Effects of types of low intensity physicalactivity (exercise/housework)in the evening on sleep in older female adults

Neural Mechanism for Hibernation
Tohru M. Takahashi1,2, Genshiro A. Sunagawa3, Shingo
Soya1, Manabu Abe4,5, Kenji Sakimura4, Takeshi
Sakurai1,6,7

Jaehoon Seol1, Simeng Zhang2, Chihiro Suzuki3,
Chihiro Kokudo4, Insung Park2, Katsuhiko Yajima5,
Makoto Satoh2, Kumpei Tokuyama2, Tomohiro Okura1

1

International Institute for Integrative Sleep Medicine (WPI-IIIS),
University of Tsukuba, Japan
2
JSPS Research Fellowship for Young Scientists, Japan
3
Laboratory for Retinal Regeneration, RIKEN Center for
Biosystems Dynamics Research, Kobe, Hyogo, Japan
4
Department of Cellular Neurobiology, Brain Research Institute,
Niigata University, Niigata
5
Department of Animal Model Development, Brain ResearchInstitute, Niigata University, Niigata
6
Faculty of Medicine, University of Tsukuba, Tsukuba, Japan
7
Life Science Center for Tsukuba Advanced Research Alliance(TARA), University of Tsukuba, Japan1

Physical Education, Health and Sport Sciences, University ofTsukuba2
International Institute for Integrative Sleep Medicine (WPIIIIS), University of Tsukuba
3
Medical Sciences, University of Tsukuba
4Sports Medicine, University of Tsukuba
5
Faculty of Health and Nutrition, Tokyo Seiei College.

Purpose: To examine different effects of exercise andhousework on sleep quality in older female adults.
Methods: The present study was a randomized and 3way crossover study. The participants were 10 healthyolderfemales,
and
sleep
was
recordedpolysomnographically. Three hours before bedtime, theparticipants were engaged in different types of lower
intensity physical activity 30 minutes (exercise orhousework) or sedentary activity (control). For the
exercise, stepping exercise was conducted at 70 beats
per minute while for the housework, housework was
conducted on about 3 METs. Subjective sleep quality
was investigated in the next morning using OSA-MA
questionnaire.
Results: The deep body temperature was significantly
elevated compared to the control after each activity (0.2, +0.5, and +0.4℃, control, housework, and exercise,respectively). There is a significant difference in sleep
latency (ANOVA P = 0.011) while there was no
significant difference among the trials (14.2, 9.9, and
4.1 min for control, housework, and exercise,respectively). An increase in stage N3 by exercise andhousework was observed during the first tertile of total
sleep time (P < 0.001). Also, stage R was significantly
longer in exercise than control and housework during
the first tertile of total sleep time (P = 0.003). The total
score of OSA-MA was significantly higher under
exercise (91.0, 88.1, and 108.6 point for control,housework, and exercise, respectively).
Conclusion: Engaging in lower intensity of exercise
and housework in the evening shows higher objective
sleep quality compared to control. However, houseworkshows lower subjective sleep quality than exercise.

Homeothermic animals, birds and mammals, consume alarge portion of their body energy for heat production tomaintain their body temperature (TB) within a narrow
range that is usually higher than the ambient
temperature. However, some mammals actively lower
TB and reduce metabolic rate to conserve energy in orderto survive food scarcity in wintertime. The
physiological state of reduced metabolic rate and TB in
endothermic animals is known as hibernation.
Hibernators return to normal condition without any
obvious tissue damage. Although numerous studies
have reported that hibernation are regulated by thecentral nervous system, neural mechanism ofhibernation has remained totally unknown. Here, we
found that neuroscientific manipulations of a novel
genetically-defined neural population in the
hypothalamus
drivealong-lasting
hypometabolic/hypothermic state (hibernation-likestate) in mice which cannot have ability to hibernate. In
this state, the mice show remarkable reductions inmetabolic rate, heartbeat, ventilation and theoretical setpoint TB. With reduced these body functions, mice also
demonstrate
that
their
thermosensory
andthermoregulatory systems remain functional, as well as
examples of hibernator. In addition, mice spontaneously
recovered from this state without obvious abnormalities,
suggesting that the induced hypometabolic state are notpassive, but actively regulated like hibernation. Ourpresent finding sheds light on the mechanism of
regulated hypometabolism, and can provide a key ofdeveloping a method to artificially induce a systemic
hibernation-like hypometabolic state in non-hibernators
including human being.
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P25

Deciphering a novel neural circuit in thecentral amygdala implicated in regulation ofsocial behavior

Decoding the sexual behavior of male miceby capturing the dopamine dynamics /Development of the automated analyzing
system for social behavior

Shingo Soya*, Katsuyasu Sakurai, Manabu Abe, Kenji
Sakimura, Takeshi Sakurai

Ai Miyasaka1,2,4, Miyabi Endo3, Ai Yoshida3, Shumpei
Yaita4 Ayano Fujinaka4, Katsuyasu Sakurai1, Hotaka
Takizawa4, Qinghua Liu1*

International Institute for Integrative Sleep Medicine (WPIIIIS), University of Tsukuba, Japan1

The regulation of interpersonal distance i.e. social space
plays a central role in social behavior, and intrusions
into personal space often evoke negative emotion insocial interactions. However, the neural mechanism
regulating interpersonal distance remains unknown.
Here, we identified a novel genetically-defined
population of neurons that express neuropeptide B/W
receptor 1 (Npbwr1 neurons) in the central nucleus of
the amygdala (CeA) as an essential component for
regulating interpersonal distance in mice. We generated
Npbwr1-iCre knocked-in mice and used for various
manipulation. Chemogenetic manipulation of Npbwr1
neurons in the CeA with excitatory and inhibitory
DREADD (hM3Dq, hM4Di) bidirectionally modulatedsocial interaction with a novel conspecific. To identify
input and output architecture of Npbwr1 neurons, wetook anterograde and retrograde tracing approach. Cellspecific infection of Rabies virus (SAD Δ G-GFP)
allowed us to visualize monosynaptic input neurons in
the CA1, BNST, NAc and VTA. Cell specific infectionof AAV carrying synaptophysin-mCherry identified thePBN, NTS and microcellular tegmentum (MiTg) as
potential postsynaptic targets. Pathway specificmanipulation of Npbwr1 neurons projecting MiTg
modulated time of social interaction with a conspecificmouse. These results suggested that Npbwr1 neurons in
the CeA projecting to the MiTg plays a crucial role inmaintenance of interpersonal distance with between
conspecifics. These findings shed light on the role ofNpwbwr1 to regulate interpersonal distance showing a
possibility to develop a new drug to treat the abnormalsociability sometimes seen in social phobia oradjustment disorder.

WPI-IIIS Liu/Sakurai Laboratory
Ph.D. Program in Humanics, University of Tsukuba
3
University of Tsukuba, Medicine group, Medical science4
Machine Intelligence & Biomedical Engineering Laboratory,
Intelligent Image Processing Group2

Sexual behavior is unsupervised innate behavior
underlying reproductive success and widely observed inmany species. In male rodents, sexual behavior iscomposed of two components: the appetitive andconsummatory behaviors. The appetitive behavior
reflects a motivational state and consists of searching
and approaching a female. The appetitive behavior leads
to the consummatory behaviors include mounting,intromission, and ejaculation. How the brain
implements a sequential but complex sexual behavior isstill unclear. Neuromodulator dopamine (DA) works as
the most prominent reward and motivation related
signals in the brain. Furthermore, DA is involved incognitive and motor tasks. We hypothesize that DA
signals in the Nucleus accumbense (NAc) modulates orcontrols the particular components of sexual behavior in
the male mouse. Establishing a causal link between DA
signaling and each male sexual behavior component, themeasuring dopamine released with high spatiotemporal
precision is necessary. Here, the in vivo fiber
photometry system is used to measure dopamine
dynamics in the NAc. Through measuring dopamine
signaling in NAc during male mouse sexual behavior by
combining fiber photometry and GRABDA, geneticallyencoded GPCR-activation-based-dopamine sensors, weobserved characteristic dopamine signaling patterns
depending on the region of NAc. During appetitive
behaviors, DA release is transiently increased when afemale mouse is introduced in a male mouse. Duringconsummatory
behaviors,
especiallyduringintromission and after ejaculation, two opposite patternsof DA release are observed: increased DA release or
suppressed DA release.
As a concurrent project, I’m developing an automatedbehavior analyzing system for identifying male sexual
behavior.
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Neural dynamics of VTA dopaminergic
neurons in female sexual behavior

Novel neural circuits for innate fear induced
defensive responses.

Kai Yashiro, Ai Miyasaka, Liu Qinghua, Katsuyasu
Sakurai, Takeshi Sakurai

Can Liu, Chia-Ying Lee, Greg Asher, Qinghua Liu,
Katsuyasu Sakurai

International Institute for Integrative Sleep Medicine (WPIIIIS), University of Tsukuba, Japan

International Institute for Integrative Sleep Medicine (WPIIIIS), University of Tsukuba, Japan

Dopamine has been thought to have a crucial role insexual behavior in both male and female animals.
However, the neural dynamics of VTA dopaminergic
neurons in a female mouse during sexual behavior
remain unclear. Here, we examined the neural activities
of VTA dopaminergic neurons in a female mouse during
sexual behavior. We used fiber photometry, whichenables us to measure the bulk of neural activities of thegenetically specific neural population with hightemporal precisions. VTA dopaminergic neurons arelabeled with a genetically encoded calcium indicator,
GCaMP6s, by applying Cre-dependent GCaMP6s
expressing AAV in Dopamine transporter (DAT)-Cre
female mice. We discovered the dramatic surge of theneural activity of VTA dopaminergic neurons in thefemale mouse immediately after male mouseejaculation. Although it is unclear the biologicalmeaning of this surge of dopaminergic neurons
activities in female mice during sexual behavior, we
speculate that this would be related to social memory or
recognition. Furthermore, the trigger of this rapid
increase in neural activities is unknown. We are thinking
several candidates; tactile or chemical input from sperm,
ultrasonic vocalization of a male mouse.

Freezing has been recognized as a typical defensive
behavior in predator related innate fear condition formany years. Besides freezing behavior, prey animals
also show significant body temperature change as thephysiological defensive strategy for survival when
encountering their predators. But its neural mechanism
is rarely studied before.
Here, we find predator odor 2MT induces potent
freezing and hypothermia in mice and identify thatexternal parabrachial nucleus (PBel) and their input toparasubthalamic nucleus (PSTh) contribute to 2MT
induced innate fear responses. Optogenetic activation of
fos expressed neurons in PBel, and their axon terminals
in PSTh with CANE system induces both freezing andhypothermia in mice. Conversely, inhibition of Vglut2
positive neurons in PBel and PSTh with DREADD
system attenuates 2MT induced freezing andhypothermia.
Moreover, we find hypothermia is regulated by the
nucleus of solitary tract (NST) projecting PSTh neurons.
Optogenetic activation of NST projecting neurons in
PSTh induces hypothermia but not freezing, which hints
that there exists another efferent brain region from PSThfor freezing regulating.
In conclusion, we identified novel neural circuits
regulating predator odor induced freezing andhypothermia responses.

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The functional role of delta-opioid system onchronic exposure to social defeat stress

Signal dynamics corresponding to value-tochoice transformation in midbrain dopamine
neurons and orbitofrontal neurons during
economic decision-making in monkeys

Akihisa Tokuda1,2, Yasuyuki Nagumo1, Hiroshi Nagase1
1

Master’s Program in Medical Sciences, University of Tsukuba,
Japan
2
International Institute for Integrative Sleep Medicine (WPI-IIIS),
University of Tsukuba, Japan

Mengxi Yun1, Takashi Kawai2, Masafumi Nejime2,
Hiroshi Yamada2, Masayuki Matsumoto2
1
2

Benzodiazepines, tricyclic antidepressants and SSRIs(Selective Serotonin Reuptake Inhibitors) are generally
used for mood or anxiety disorder. However, these drugs
need a long-term taking to obtain therapeutic effects, oroften induce adverse effects. These evidences suggest
the necessity for new attractive therapeutic targets inmood or anxiety disorder. Delta-opioid neurons exist in
cortical and limbic structures, and produce uniqueemotional function in the brain. Here, we report deltaopioid neural function on exposure to chronic social
defeat stress and delta-opioid receptor (DOR) agonistinduced therapeutic effectiveness on social defeat stressmodel. Chronic exposure to social defeat stress induced
the depression-like behaviors characterized by anxiety,anhedonia and social avoidance behavior in normal
mice. These behavioral changes following chronicsocial stress were recovered by repeated treatment of
KNT-127, a selective DOR agonist. This recovery wasobserved from 3 days after KNT-127 treatment. In
contrast to that, pre-treatment with NTI, a selective
DOR antagonist, induced the exacerbation ofdepression-like behaviors following social stress inmice from the beginning of stress exposure, compared
to pre-treatment with saline. These results suggest deltaopioid neural system plays a key role in stress defense
mechanism in the brain and DOR agonist is expected to
be a more effective drug than existing drugs.

Biomed. Science, Grad Sch. of Comprehensive Human Sci.
Fac. of Med., Univ. of Tsukuba, Tsukuba, Japan

In economic decision-making, individuals first evaluate
an option, and then decide to choose or not to choose theoption based on its value. Although the cortical system,
especially the orbitofrontal cortex (OFC) has beenthought as the neural substrate of the transformation
process from option’s value information into animal’s
choice command, the whole picture of neural network
remains unclear. Here we investigated the role of
midbrain dopamine (DA) neuron, a key region forvalue-processing, and compared their role with that of
the OFC. We recorded single-unit activity from DA and
OFC neurons in monkeys performing an economic
decision-making task. We found that both DA and OFC
neurons represented diverse signals related not only tothe option's value but also to the animal's choice; someneurons represented the value of the offered option,some represented whether the animal would choose or
not choose the option, and some represented the valueof the option only when the option was chosen by themonkey -we therefore called this activity pattern as
choice-dependent value signal that was influenced byboth value and choice. We next analyzed the time courseof these signals and found that the order of signal
representations in both regions corresponded to thevalue-to-choice transformation. Shortly after the onsetof the option, the value signal rapidly appeared, which
was followed by the choice-dependent value signal. The
choice signal arose at last. Our findings provide
evidence that not only cortical system but also thesubcortical DA system may participate in value-tochoice transformation.

15