# Presentation: 238080

## Experience-Dependent Alterations in

**Experience-Dependent Alterations in**
** ****the Function of Brain Reward Systems:**
**The Role of CREB**

**William A. Carlezon, Jr., Ph.D.**
**Behavioral Genetics Laboratory**
**Department of Psychiatry**
**Harvard Medical School**
**McLean Hospital**

**Notes:**
Talk today about research in my lab without showing any of our actual data

Instead, I am going to tell you about what we do, how we do it, and why we do it that way.

I hope by the end that you will see that our approach is different than most others use, and that this is a good thing.

## Addiction and Depressive Disorders

**Addiction and Depressive Disorders**

- •  **High rates of co-morbidity in both men and women**
**   ****(Kessler et al., 1997).**

- •  **Causal relationships can be difficult to establish.**

- •  **Addictive behaviors and depressive disorders have**
**   ****common triggers (e.g., stress).**

- •  **Studies of addiction may give us insights on depressive **
**   ****disorders, and vice-versa.**

## Working Hypothesis

**Addiction and Depressive Disorders**
**Involve Brain Reward Systems**

## Mood in Rodents?

## Mood in Rodents?

**Mood in Rodents?**

**Sad**

**Grumpy**

**Happy**

***Anhedonia***

***Reward***

***Dysphoria***

**Worried**

***Anxiety***

**Hopeless**

***Despair***

**Listless**

***Hypoactivity***

## Mood in Rodents

**Mood in Rodents**

- •  **reward** (feelings of pleasure)
- •  **anhedonia** (reduced ability to experience rewarding
-    things as rewarding)
- •  **dysphoria** (feelings of discomfort)
- •  **despair** (feelings of giving up)
- •  **anxiety **(feelings of apprehension, fear)
- •  **hypoactivity** (less active than normal)

## Overview

**Overview**

**1.  CREB**

**2.  Functional role of CREB in the NAc**

**3.  CREB and dynorphin**

**4.  Kappa antagonists and agonists **

## Overview

**Overview**

**1.  CREB**

**2.  Functional role of CREB in the NAc**

**3.  CREB and dynorphin**

**4.  Kappa antagonists and agonists**** **

## CREB (cAMP Response Element Binding Protein)

## Addictive Drugs Regulate CREB in the NAc

**Addictive Drugs Regulate CREB in the NAc**

**CREB phosphorylation**

**Turgeon et al., 1997**

## Drug Regulation of CREB

**Drug Regulation of CREB**

**Are there behavioral correlates of CREB**
**activity in the NAc?**
**	****•  ****Drug tolerance? **
**	****•  ****Drug sensitization?**

## Elevation of CREB with Engineered Viruses

**NAc**

**VTA**

**Elevation of CREB with Engineered Viruses**

## Elevation of CREB with Engineered Viruses

**CREB**

**NAc**

**VTA**

## Blockade of CREB Function By Overexpression

**of mCREB with Engineered Viruses**

**mCREB**

**NAc**

**VTA**

## pHSV PrpUC

**pHSV PrpUC**

**cDNA (e.g. CREB, **
**mCREB, LacZ)**

**HSV-1**
**c Region**

**pBR 322**

**a**
**HSV-1**
**packaging**
**site**

**SV-40  **
**polyadenylation**
**site**

**Amp**

**  ****Col**
** ****E1**
**ori**

**ori****s**

**IE 4/5 Promoter**

**(Neve Lab)**

## Gene Transfer Using Viral Vectors

**Gene Transfer Using Viral Vectors**

-   **Increased Protein Expression****	****           Minimal Damage**

## Overview

**Overview**

**1.  CREB**

**2.  Functional role of CREB in the NAc**

**3.  CREB and dynorphin**

**4.  Kappa antagonists and agonists**** **

## Measurement of Drug Reward (and Aversion)

## Effect of CREB Over Cocaine Doses

**Change in Time Spent in Drug Side (sec)**

***Carlezon et al., 1998***

## Intracranial Self-Stimulation (ICSS)

**Intracranial Self-Stimulation (ICSS)**
**in mCREB transgenic mice**

## ICSS Thresholds

**ICSS Thresholds**

**100%**

## Intracranial Self-Stimulation (ICSS)

**in mCREB transgenic mice**

## Forced Swim Test (FST)

***Swimming***

**Immobile**

## Elevated CREB Increases Immobility in the FST

**Time (sec)**

**Beam Breaks (1 hr)**

**Latency to Immobility**

**Locomotor Activity**

**CREB-On = More Depressive-like Signs**

**CREB-Off = Fewer Depressive-like Signs**

## Forced Swim Test (FST)

***Swimming***

**Immobile**

## Forced Swimming Activates CREB in the NAc 

**Forced Swimming Activates CREB in the NAc**** **

**Induction (swim / no swim)**

**CREB in the NAc normally plays a role in reacting to stress**

***Pliakas et al., 2001***

## Biology & Behavior

***Drugs***
***Stress***

***CREB***
***In NAc***

***Depressive***
***Behavior***

**CREB Target Genes**

## CREB (cAMP Response Element Binding Protein)

## Overview

**Overview**

**1.  CREB**

**2.  Functional role of CREB in the NAc**

**3.  CREB and dynorphin**

**4.  Kappa antagonists and agonists**** **

## Dynorphin

**Dynorphin**

**•  ****It has CRE sites in its promoter (Douglass et al., 1994)**

**•  ****It is regulated by drugs of abuse (Hurd and Herkenham,**
**   ****1993; Daunais et al., 1993; Cole et al., 1995)**

**•  ****Kappa agonists are aversive rats (Bals-Kubik et al., 1993)**

**•  ****Kappa agonists are aversive in people (Pfeiffer et al.,**
**    ****1986)**

**•  ****It is a kappa opioid receptor ligand (Chavkin et al., 1982)**

## CREB Increases Dynorphin mRNA

**in the NAc**

**CREB-On =**
**More Dynorphin**

**CREB-Off =**
**Less Dynorphin**

***Carlezon et al., 1998***

## Stress Increases Dynorphin mRNA

**in the NAc**

**Stress = More Dynorphin**

***Chartoff et al., unpublished***

## Biology & Behavior

***Drugs***
***Stress***

***CREB***
***In NAc***

***Depressive***
***Behaviors***

## Biology & Behavior

***Drugs***
***Stress***

***CREB***
***In NAc***

***Depressive***
***Behaviors***

***dynorphin***

**Clinical Implications**

## Overview

**Overview**

**1.  CREB**

**2.  Functional role of CREB in the NAc**

**3.  CREB and dynorphin**

**4.  Kappa antagonists and agonists **

## U-69593 (kappa agonist) increases immobility

**and decreases swimming **

***Mague et al., 2003***

## ANTI (IP) decreases immobility

**and increases climbing **

***Mague et al., 2003***

****ANTI: 5’-acetamidinoethylnaltrindole***

## Proposed Circuitry

**Proposed Circuitry**

**Carlezon et al., 2000**

## X

**Proposed Circuitry**

**Carlezon et al., 2000**

**X**

**X**

## Proposed Circuitry

**Proposed Circuitry**

**Carlezon et al., 2000**

**X**

**X**

## Kappa-selective agonist

- O

- O

- O

- O

- O

- O

- O

- O

**Salvinorin A**

*(kappa agonist)*

***(Roth et al., 2002 )***

## Salvinorin A

**Salvinorin A**

**Microdialysis (NAc)**

***(Carlezon et al., 2005)***

## Abuse Liability of Kappa Antagonists?

**Abuse Liability of Kappa Antagonists?**

## Effect of kappa ligands on ICSS

**Effect of kappa ligands on ICSS**

## Effect of kappa ligands on ICSS

**Effect of kappa ligands on ICSS**

## Effect of kappa ligands on ICSS

**Effect of kappa ligands on ICSS**

## Effect of kappa ligands on ICSS

**Effect of kappa ligands on ICSS**

## Medicinal Chemistry Program

- O

- O

- O

- O

- O

- O

- O

- O

**Salvinorin A**

*(kappa agonist)*

**Furan-ring derivatives**

**Lactone-ring derivatives**

**C2 derivatives**

**C4 derivatives**

## Does CREB in the NAc Regulate Mood?

**Does CREB in the NAc Regulate Mood?**

***Carlezon & Konradi, 2004***

## Acknowledgements

**Acknowledgements**

_**Carlezon Lab**_

**Andrea Pliakas**
**Andreas Arvanitogiannis**
**Rebecca Ralph-Williams**
**Sonia Guediche**
**Steve Mague**
**Brian Gilliss**
**Elena Chartoff**
**Hilarie Tomasiewicz**
**Mark Todtenkopf**
**Edward Meloni**
**Alexandra Jackson**
**Jacqueline Marcus**
**Maria Papadopoulou**
**Jennifer DiNieri**
**Katie Famous**
**Andrew Smith**
**Jessica Pohlman**
**Aram Parsegian**
**Melissa Stoos**
**Lyle Gerety**
**Tracie Paine**
**Dost Ongur**
**Allison Knoll**
**Matthew Barhight**
**Antonia Dow**

_**Funding Sources**_
**NIDA, NIMH, NARSAD**

_**McLean Labs**_
**Bruce Cohen, Christine Konradi**
**Rachael Neve, Susan Andersen**

## Special Acknowledgements

**Special Acknowledgements**

**Waletzky Family**

**Society for Neuroscience**

## Kappa Antagonists have Antidepressant Effects

**Kappa Antagonists have Antidepressant Effects**

**Time (sec)**

**Beam Breaks (1 hr)**

**Locomotor Activity**

**Latency to Immobility**

## “CREB-On”

- CRE

*“**CREB-On”*

*“**CREB-Off”*

## Kappa-selective agonist

**Salvinorin A**

*(kappa agonist)*

*Salvia Divinorum*

## Salvinorin A

***(Carlezon et al., JPET, in press)***

**Salvinorin A**

## Salvinorin A

**Salvinorin A**

***(Carlezon et al., JPET, in press)***

## Salvinorin A

**Salvinorin A**

**Microdialysis (NAc)**

***(Carlezon et al., JPET, in press)***