Graduate School ETD Form 9 (Revised 12/07) PURDUE UNIVERSITY GRADUATE SCHOOL Thesis/Dissertation Acceptance This is to certify that the thesis/dissertation prepared By Scott M Oster Entitled Chronic Ethanol Drinking by Alcohol-preferring Rats Increases the Sensitivity of the Mesolimbic Dopamine System to the Reinforcing and Stimulating Effects of Cocaine Doctor of Philosophy For the degree of Is approved by the final examining committee: James M Murphy Cristine L Czachowski Chair Zachary A Rodd Charles R Goodlett Kimberly P Kinzig To the best of my knowledge and as understood by the student in the Research Integrity and Copyright Disclaimer (Graduate School Form 20), this thesis/dissertation adheres to the provisions of Purdue University’s “Policy on Integrity in Research” and the use of copyrighted material. Approved by Major Professor(s): ____________________________________James M Murphy ____________________________________ Approved by: John T Hazer 02/10/2012 Head of the Graduate Program Date Graduate School Form 20 (Revised 9/10) PURDUE UNIVERSITY GRADUATE SCHOOL Research Integrity and Copyright Disclaimer Title of Thesis/Dissertation: Chronic Ethanol Drinking by Alcohol-preferring Rats Increases the Sensitivity of the Mesolimbic Dopamine System to the Reinforcing and Stimulating Effects of Cocaine For the degree of DoctorChoose ofyour Philosophy degree I certify that in the preparation of this thesis, I have observed the provisions of Purdue University Executive Memorandum No. C-22, September 6, 1991, Policy on Integrity in Research.* Further, I certify that this work is free of plagiarism and all materials appearing in this thesis/dissertation have been properly quoted and attributed. I certify that all copyrighted material incorporated into this thesis/dissertation is in compliance with the United States’ copyright law and that I have received written permission from the copyright owners for my use of their work, which is beyond the scope of the law. I agree to indemnify and save harmless Purdue University from any and all claims that may be asserted or that may arise from any copyright violation. Scott M Oster ______________________________________ Printed Name and Signature of Candidate 02/10/2012 ______________________________________ Date (month/day/year) *Located at http://www.purdue.edu/policies/pages/teach_res_outreach/c_22.html CHRONIC ETHANOL DRINKING BY ALCOHOL-PREFERRING RATS INCREASES THE SENSITIVITY OF THE MESOLIMBIC DOPAMINE SYSTEM TO THE REINFORCING AND STIMULATING EFFECTS OF COCAINE A Dissertation Submitted to the Faculty of Purdue University by Scott M. Oster In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy May 2012 Purdue University Indianapolis, Indiana ii For Ty, Xaven, and Daxen. iii ACKNOWLEDGMENTS I would like to express gratitude to my advisor, Dr. James Murphy, and doctoral committee: Dr. Zachary Rodd, Dr. Charles Goodlett, Dr. Cristine Czachowski, and Dr. Kimberly Kinzig. These committee members are directly responsible for the achievement of a doctorate degree through their continued encouragement and support. I am also grateful to Dr. William McBride for his for the unwavering guidance and support, as well as for the continued use of laboratory facilities and experimental equipment. In addition, this dissertation would not have been possible without the technical support, insight, and constructive feedback of Dr. Eric Engleman, Dr. Jamie Toalston, Dr. Gerald Deehan, Dr. Sheketha Hauser, Dr. Zhengmeng Ding, and Dr. Simon Katner. Lastly, I would like to thank Tylene Pommer, Victoria McQueen, Curtis Bard, Erin Larrabee, Freida Cormack, Mary Ann Hindman, and Heather Sissons for essential technical and administrative assistance. This project was supported in part by NIAAA Grants AA10717, AA12262, and AA07611, as well as Ruth L. Kirschstein National Research Service Award Individual Predoctoral Fellowship Grant AA016429. iv TABLE OF CONTENTS Page LIST OF FIGURES .............................................................................................. vii LIST OF ABBREVIATIONS .................................................................................. ix ABSTRACT..........................................................................................................xiii 1. INTRODUCTION ...............................................................................................1 Addiction and Polydrug Use.............................................................................1 The Role of Mesocorticolimbic Dopamine........................................................4 Behavioral and Neurobiological Assays...........................................................7 Mesoaccumbens Circuit: Connectivity and Heterogeneity.............................10 Ventral Tegmental Area: Connectivity ......................................................10 Nucleus Accumbens: Connectivity............................................................12 Ventral Tegmental Area: Regional Heterogeneity ....................................14 Nucleus Accumbens: Regional Heterogeneity..........................................15 Drugs of Abuse Produce Alterations in Mesocorticolimbic Functioning .........17 Acute Reinforcing and Dopamine Neuronal Stimulating Effects of Cocaine within the Mesoaccumbens Circuit...................................................18 Chronic Ethanol Exposure: Effects on Neurobiology and Behavior ...............20 Genetic Factors Associated with Drug and Alcohol Abuse ............................21 Objectives and Experimental Rationale .........................................................25 2. METHODS.......................................................................................................27 Animals...........................................................................................................27 Drinking Procedures.......................................................................................28 Chemical Agents ............................................................................................29 Histology.........................................................................................................30 Sensitization of Cocaine Reinforcement within the Nucleus Accumbens Shell Following Chronic Ethanol Drinking...................................30 Animal Preparation ...................................................................................30 Intracranial Self-administration Apparatus................................................31 General Test Condition.............................................................................32 Experimental Groups ................................................................................34 Statistical Analysis ....................................................................................34 Persistent Alteration of Posterior Ventral Tegmental Area Dopamine Neurons to the Stimulating Effects of Cocaine Following Chronic Ethanol Drinking.............................................................................................35 Animal Preparation ...................................................................................35 General Test Condition.............................................................................36 v Page Experimental Groups ................................................................................37 Microdialysis Sample Analysis..................................................................37 Statistical Analysis ....................................................................................38 3. RESULTS ........................................................................................................39 Sensitization of Cocaine Reinforcement within the Nucleus Accumbens Shell Following Chronic Ethanol Drinking ......................................................39 Histology ...................................................................................................39 Ethanol Intake for Chronic Ethanol-drinking Rats.....................................40 Acquisition of Cocaine Self-infusion: Sessions 1-4...................................41 Extinction of Cocaine Self-infusion: Sessions 5-7.....................................44 Reinstatement of Cocaine Self-infusion: Session 8..................................45 Lever Responding of Ethanol-naïve Rats .................................................49 Lever Responding of Ethanol-drinking Rats .............................................54 Infusions and Lever Responding of Anatomical Controls .........................56 Persistent Alteration of Posterior Ventral Tegmental Area Dopamine Neurons to the Stimulating Effects of Cocaine Following Chronic Ethanol Drinking.............................................................................................59 Histology ...................................................................................................59 Ethanol Intake for Ethanol-drinking and Ethanol-abstinent Rats ..............61 Temporal Pattern of the Stimulating Effects of Cocaine on Posterior Ventral Tegmental Area Dopamine Neurons............................................61 Nucleus Accumbens Shell Dopaminergic Response After Injection of Artificial Cerebrospinal Fluid or 0.25 mM Cocaine into the Posterior Ventral Tegmental Area............................................................................63 Nucleus Accumbens Shell Dopaminergic Response After Injection of 0.5 mM Cocaine into the Posterior