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Behavioral and Molecular Analysis of Individual Variation in Ethanol Drinking Virginia Commonwealth University VCU Scholars Compass Theses and Dissertations Graduate School 2009 Behavioral and Molecular Analysis of Individual Variation in Ethanol Drinking Jennifer Wolstenholme Virginia Commonwealth University Follow this and additional works at: https://scholarscompass.vcu.edu/etd Part of the Medical Pharmacology Commons © The Author Downloaded from https://scholarscompass.vcu.edu/etd/1874 This Dissertation is brought to you for free and open access by the Graduate School at VCU Scholars Compass. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of VCU Scholars Compass. For more information, please contact [email protected]. School of Medicine Virginia Commonwealth University This is to certify that the dissertation prepared by Jennifer T. Wolstenholme entitled BEHAVIORAL AND MOLECULAR ANALYSIS OF INDIVIDUAL VARIATION IN ETHANOL DRINKING has been approved by his or her committee as satisfactory completion of the dissertation requirement for the degree of Doctor of Philosophy Michael F. Miles, MD, PhD John W. Bigbee, PhD, School of Medicine Michael C. Neale, PhD, School of Medicine Keith L. Shelton, PhD, School of Medicine Laura J. Sim-Selley, PhD, School of Medicine William L. Dewey, PhD, Interim Chair of Pharmacology and Toxicology Jerome F. Strauss, III, MD, PhD, Dean, School of Medicine Dr. F. Douglas Boudinot, Dean of the Graduate School July 23, 2009 © Jennifer Theresa Wolstenholme 2009 All Rights Reserved BEHAVIORAL AND MOLECULAR ANALYSIS OF INDIVIDUAL VARIATION IN ETHANOL DRINKING A Dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy at Virginia Commonwealth University. By JENNIFER THERESA WOLSTENHOLME Bachelor of Arts, University of Virginia, 1996 Director: MICHAEL F. MILES, MD, PhD Professor, Departments of Pharmacology/Toxicology and Neurology Virginia Commonwealth University Richmond, Virginia July, 2009 Acknowledgements I would like to thank the numerous people who have supported me throughout these years of intense study, work and writing. Many thanks to my family whom have facilitated the dedication needed to complete this degree. I have been incredibly blessed to have three extraordinary children, Kyra, Omar, Jr. and Haley, who constantly brighten the days and ground me in the proper perspective. There are not enough words to describe my intense appreciation to Omar, without whom this never would have been accomplished. Thank you, Omar, for your patience, support, encouragement and love throughout these years. I would also like to thank the past and present members of Dr. Miles’ laboratory, Alex, Rob, Nate, and Sean for your help with dissections, Shauna and Jon for help with the everlasting drinking experiments, and for your support and friendship. Also, my committee members, Dr. John Bigbee, Dr. Mike Neale, Dr. Keith Shelton and Dr. Laura Sim-Selley as well as my consultant, Dr. Klaus Miczek have been wonderful academic and creative resources while working on this project. Finally, many thanks to Dr. Mike Miles for your years of teaching, encouragement, and support; not to mention, trust, allowing me to embark on this project. ii Table of Contents Page Acknowledgements ............................................................................................................. ii List of Tables ..................................................................................................................... vi List of Figures ................................................................................................................... vii Abbreviations .......................................................................................................................x Abstract ............................................................................................................................ xiii Chapter 1 Introduction ......................................................................................................1 2 Background and Significance .........................................................................4 Individual differences in ethanol drinking ....................................................4 Neurobiology of stress ...................................................................................8 Animal models of social stress ....................................................................11 Neurobiology of social stress in rodents .....................................................14 Neural pathways influencing stress and ethanol drinking ...........................18 Behavioral effects of stress and drugs of abuse ..........................................19 Role of stress in excessive drinking and alcohol abuse ...............................20 Social stress and ethanol drinking behaviors ..............................................22 Application of DNA microarrays to ethanol-related behaviors ..................25 iii 3 Characterization of Individual Variation of Ethanol Drinking in C57 Mice .............................................................................................................27 Introduction .................................................................................................27 Methods .......................................................................................................29 Results .........................................................................................................32 Discussion ...................................................................................................39 4 Social Stress and its Effects on Ethanol Drinking Behavior ......................44 Introduction .................................................................................................44 Methods .......................................................................................................47 Results .........................................................................................................53 Discussion ...................................................................................................69 5 Anxiety-Like Behaviors in Ethanol Drinking and Social Defeat ...............78 Introduction .................................................................................................78 Methods .......................................................................................................81 Results .........................................................................................................84 Discussion ...................................................................................................92 6 Molecular Factors Contributing to Individual Variation of Ethanol Drinking and Response to Social Stress ................................................101 Introduction ...............................................................................................101 Methods .....................................................................................................103 iv Results .......................................................................................................110 Discussion .................................................................................................134 7 Conclusions and Future Directions ............................................................159 Literature Cited ................................................................................................................166 Vita ...................................................................................................................................189 v List of Tables Page Table 1: Effects of Stress on Ethanol Drinking in Rodent Models....................................23 Table 2: Correlation of Initial Ethanol Intake versus Subsequent Rounds of Drinking Following Deprivation. ........................................................................................34 Table 3: Social Rank and Ethanol Intake. ..........................................................................57 Table 4: Basal Anxiety Phenotypes Versus Anxiety Phenotypes After Ethanol Drinking. ...............................................................................................................87 Table 5: Correlation of Basal Anxiety-Like Behavior with Ethanol Intake .....................89 Table 6: Over-Represented Gene Categories Significantly Correlated to Ethanol Drinking in C57BL/6NCrl Mice. ......................................................................................149 Table 7: Genes Involved in Chromatin Remodeling Identified in Nucleus Accumbens of Ethanol Drinking Mice. .....................................................................................153 Table 8: Genes from Nucleus Accumbens of Ethanol Drinking Mice Involved in Synaptic Vesicle Formation and Recycling. .....................................................154 Table 9: Over-Represented Gene Categories from Principle Component Analysis in Ethanol Drinking Mice. .....................................................................................155 Table 10: Over-Represented Gene Categories Significant for the Interaction of Ethanol Drinking and Social Defeat in the NAc of C57BL/6NCrl Mice. ......................158 vi List of Figures Page Figure 1: Stress Responsive and Ethanol Responsive Pathways in the Brain. ..................10 Figure 2: Neural Circuits for Brief Social Defeat Stress and Continuous Subordination Stress. ........................................................................................16
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