Jacob P Waletzky Memorial Lecture ExperienceDependent Alterations in

Jacob P. Waletzky Memorial Lecture 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 Mc. Lean Hospital

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? Happy Hopeless Grumpy Listless Sad Worried

Mood in Rodents? Happy Reward Hopeless Despair Grumpy Dysphoria Listless Hypoactivity Sad Anhedonia Worried Anxiety

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 1. CREB 2. Functional role of CREB in the NAc 3. CREB and dynorphin 4. Kappa antagonists and agonists

CREB (c. AMP Response Element Binding Protein)

Addictive Drugs Regulate CREB in the NAc CREB phosphorylation Turgeon et al. , 1997

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

Elevation of CREB with Engineered Viruses CREB NAc VTA

Blockade of CREB Function By Overexpression of m. CREB with Engineered Viruses m. CREB NAc VTA

p. BR 322 Amp a HSV-1 packaging site SV-40 polyadenylation site Col E 1 ori p. HSV Prp. UC oris HSV-1 c Region (Neve Lab) IE 4/5 Promoter c. DNA (e. g. CREB, m. CREB, Lac. Z)

Gene Transfer Using Viral Vectors Increased Protein Expression Minimal Damage

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)

Change in Time Spent in Drug Side (sec) Effect of CREB Over Cocaine Doses Anhedonia Dysphoria Carlezon et al. , 1998

Intracranial Self-Stimulation (ICSS) in m. CREB transgenic mice

ICSS Thresholds Aversive/Prodepressants (drug withdrawal, lithium, neuroleptics) 100% Drugs of Abuse (amphetamine, cocaine, opiates, PCP, nicotine)

Intracranial Self-Stimulation (ICSS) in m. CREB transgenic mice

Forced Swim Test (FST) Swimming Immobile

Elevated CREB Increases Immobility in the FST Locomotor Activity Beam Breaks (1 hr) Time (sec) Latency to Immobility CREB-On = More Depressive-like Signs CREB-Off = Fewer Depressive-like Signs

Forced Swim Test (FST) Swimming Immobile

Induction (swim / no swim) Forced Swimming Activates CREB in the NAc normally plays a role in reacting to stress Pliakas et al. , 2001

Biology & Behavior Drugs Stress CREB In NAc CREB Target Genes Depressive Behavior

CREB (c. AMP Response Element Binding Protein)

Overview 1. CREB 2. Functional role of CREB in the NAc 3. CREB and dynorphin 4. Kappa antagonists and agonists

Dynorphin • It is a kappa opioid receptor ligand (Chavkin et al. , 1982) • 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 in people (Pfeiffer et al. , 1986) • Kappa agonists are aversive rats (Bals-Kubik et al. , 1993)

CREB Increases Dynorphin m. RNA in the NAc CREB-On = More Dynorphin CREB-Off = Less Dynorphin Carlezon et al. , 1998

Stress Increases Dynorphin m. RNA 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 dynorphin Clinical Implications Depressive Behaviors

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 *ANTI: 5’-acetamidinoethylnaltrindole Mague et al. , 2003

Proposed Circuitry Carlezon et al. , 2000

Proposed Circuitry X X Carlezon et al. , 2000

Kappa-selective agonist O O O O Salvinorin A (kappa agonist) (Roth et al. , 2002 )

Salvinorin A Microdialysis (NAc) (Carlezon et al. , 2005)

Abuse Liability of Kappa Antagonists?

Effect of kappa ligands on ICSS

Medicinal Chemistry Program O O O C 2 derivatives O Furan-ring derivatives Lactone-ring derivatives O O C 4 derivatives Salvinorin A (kappa agonist)

Does CREB in the NAc Regulate Mood? Carlezon & Konradi, 2004

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 Di. Nieri Katie Famous Andrew Smith Jessica Pohlman Aram Parsegian Melissa Stoos Lyle Gerety Tracie Paine Dost Ongur Allison Knoll Matthew Barhight Antonia Dow Mc. Lean Labs Funding Sources Bruce Cohen, Christine Konradi Rachael Neve, Susan Andersen NIDA, NIMH, NARSAD

Special Acknowledgements Waletzky Family Society for Neuroscience

Kappa Antagonists have Antidepressant Effects Locomotor Activity Beam Breaks (1 hr) Time (sec) Latency to Immobility

CBP/p 300 HAT A CRE “CREB-On” RNA Pol-II CBP/p 300 P CREB m. CREB X CRE “CREB-Off” RNA Pol-II

Kappa-selective agonist O O O O Salvinorin A (kappa agonist) Salvia Divinorum

Salvinorin A FST (Carlezon et al. , JPET, in press)

Salvinorin A ICSS (Carlezon et al. , JPET, in press)

Salvinorin A Microdialysis (NAc) (Carlezon et al. , JPET, in press)