EFFECTS OF CHRONIC METHYLPHENIDATE ON DOPAMINE/SEROTONIN INTERACTIONS IN THE MESOLIMBIC DA SYSTEM OF THE MOUSE BY BETHANY R. BROOKSHIRE A Dissertation Submitted to the Graduate Faculty of WAKE FOREST UNIVERSITY GRADUATE SCHOOL OF ARTS AND SCIENCES in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY Physiology and Pharmacology May 2010 Winston-Salem, North Carolina Approved by: Sara R. Jones, Ph.D., Advisor ___________________________________ Examining Committee: Dwayne W. Godwin, Ph.D., Chairman ____________________________________ David C. S. Roberts, Ph.D. ____________________________________ Brian McCool, Ph.D. ____________________________________ Jeff L. Weiner, Ph.D. ____________________________________ Paul W. Czoty, Ph.D. ____________________________________ i ACKNOWLEDGEMENTS Firstly, I would like to express my gratitude to my advisor, Sara R. Jones, PhD. Working in her lab has been both a wonderful educational experience as well as a pleasure. She had always offered guidance, support, and numerous professional opportunities. I am proud to have been trained in her laboratory. I would also like to acknowledge the excellent guidance of my committee: Drs. Godwin, Roberts, McCool, Weiner, and Czoty. They have never hesitated to offer mentorship and advice, and have been instrumental in the development of my work. Additionally, I would like to express my gratitude to Dr. Mary Lou Voytko, who has never hesitated to give me sound advice and mentorship. I would like to thank the members of the Jones Lab, past and present. Drs. Tiffany Mathews and Yolanda Mateo for their fantastic support and training, Drs Rodrigo Espana and Mark Ferris, for their fantastic professional guidance, and Joanne Konstantopolous and Jason Locke, for their constant technical assistance and support. I would also like to acknowledge Dr Kim Huggins, Dr. Erik Oleson, Jordan Yorgason, Erin Calipari, and James Melchior, they have always helped to make the lab an enjoyable experience. I would like to acknowledge the support of the Department of Physiology and Pharmacology and the Neuroscience Program. Their members have always been a willing source of advice. Special thanks to Stephanie Williard, Dr. Keri Chiodo, Dr. Anushka Goonadwardena, Tamara Spence, Dr. John Burkhardt, Kimberly Bernosky, and Dr. Greg Tinkler for their unwavering support and friendship. ii I cannot express enough thanks to my family and friends. My father, Dr. Bob Brookshire, my mother, Cathy Brookshire, and my brother, David Brookshire, who have been endlessly supportive and caring, and never stop making me laugh. I also want to recognize Mr. Vincent Piattelli, who is always a source of caring, support, sound advice, and perspective. My friends both in Winston Salem and everywhere else, who are always there to help motivate me when I need it, and who have helped to shape me into who I am, personally and professionally. I would particularly like to recognize Dr. Paul Heideman, my undergraduate mentor, who always encouraged me scholastically, and whose excellent lectures in Animal Physiology put me on the path toward graduate school. iii TABLE OF CONTENTS Page LIST OF ABBREVIATIONS…………………………………………..…………… vi LIST OF TABLES AND FIGURES ……………………….………………………. ix ABSTRACT …………………………………………………………………………. xiv Chapter I. INTRODUCTION. SECTION I: EFFECTS OF ACUTE AND CHRONIC METHYLPHENIDATE ON THE NEUROBIOLOGY AND BEHAVIOR OF RATS …………………………………………. 1 SECTION II: EFFECTS OF SPECIFIC 5-HT RECEPTOR SUBTYPES ON ACTIVATION OF DOPAMINE SIGNALING AND PSYCHOSTIMULANT RESPONSES IN THE VENTRAL TEGMENTAL AREA/NUCLEUS ACCUMBENS 47 PATHWAY……………………………………………………… II. CHRONIC METHYLPHENIDATE TREATMENT REVERSES NEGATIVE SEROTONIN EFFECTS OF DRUG REWARD: A ROLE FOR 5-HT1B RECEPTORS …………………………….. 116 Submitted to Neuropsychopharmacology, January, 2010. III. SENSITIZATION OF 5-HT1A and 1B RECEPTORS IN THE VENTRAL TEGMENTAL AREA FOLLOWING TREATMENT WITH CHRONIC METHYLPHENIDATE ……. 155 In preparation for submission to Neuropsychopharmacology, May, 2010. IV. CHRONIC MPH ADMINISTRATION IN MICE PRODUCES DEPRESSIVE-LIKE EFFECTS AND ALTERED BEHAVIORAL RESPONSES TO ANTIDEPRESSANTS …… 200 In preparation for submission to Pharmacology, Biochemistry, and Behavior, March, 2010. iv V. TEMPORARY EFFECTS OF CHRONIC MPH ON DA/5-HT INTERACTIONS IN MICE: EVIDENCE FOR 5-HT INVOLVEMENT IN EARLY BEHAVIORAL SENSITIZATION ……………………………………………….. 229 In preparation for submission to Psychopharmacology, March, 2010. VI. DIRECT AND INDIRECT 5-HT RECEPTOR AGONISTS PRODUCE GENDER-SPECIFIC EFFECTS ON LOCOMOTOR AND VERTICAL ACTIVITY IN C57 BL/6J MICE ……………………………………………………………. 259 Published in Pharmacology, Biochemistry, and Behavior, August, 2009. VII. SUMMARY AND DISCUSSION ……………………………… 323 VIII. SCHOLASTIC VITA …………………………………………… 348 IX. APPENDIX I: ETHANOL-INDUCED HYPERACTIVITY IS ASSOCIATED WITH HYPODOPAMINERGIA IN THE 22- TNJ ENU-MUTATED MOUSE ……………………………….. 354 Published in Alcohol, September 2009 v LIST OF ABBREVIATIONS [DA]p 5-HT: 5-Hydroxytryptamine, Serotonin 5-HT1A, 1A: Serotonin 1A receptor subtype 5-HT1B, 1B: Serotonin 1B receptor subtype 5-HT1B KO: Serrotonin 1B knockout mouse 5-HT1D, 1D: Serotonin 1D receptor subtype 5-HT2A, 2A: Serotonin 2A receptor subtype 5-HT2C, 2C: Serotonin 2C receptor subtype 5-HT6, 6: Serotonin 6 receptor subtype 8-OH-DPAT: R(+)-8-hydroxy-2-(di-n-propylamino)-tetralin ADHD: Attention-deficit/hyperactivity disorder ANOVA: Analysis of variance BCA: Bicinchoninic Acid CGRP: Calcitonin gene-related peptide CNS: Central nervous system CPBG: 1-(m-chlorophenyl)-biguanide CPP: Conditioned place preference CPu: Caudate putamen DA: Dopamine DAp: Dopamine release per stimulus DAT: Dopamine transporter DAT-KO: Dopamine transporter knock-out mouse vi DHBA: 3,4-Dihydroxybenzylamine DOI: 2,5-Dimethoxy-4-iodoamphetamine DRD4: Dopamine D4 receptor subtype DRN: Dorsal raphe nucleus ENU: N-ethyl-N-nitrosourea ERK-2: Extracellular signal-regulated kinase 2 GABA: γ-Aminobutyric acid GBL: Gamma-butyrolactone GR 127, 935: N-[4-methoxy3-(4-methyl-1-piperazinyl)phenyl]-2'-methyl-4'-(5me thyl- 1,2,4-oxadiozol-3-yl)-biphenyl-4-carboxamide GR 55562: 3-(3-dimethylaminopropyl)-4-hydroxy-N-(4-pyridin-4-ylphenyl)- benzamide hydrochloride HPLC: High performance liquid chromatography L-DOPA: L-3,4-dihydroxyphenylalanine mCPP: meta-Chlorophenylpiperazine MDL 100,907: (R)-(+)-α-(2,3-Dimethoxyphenyl)-1-[2-(4-fluorophenyl)ethyl]-4- piperidinemethanol MDMA: 3,4-Methylenedioxymethamphetamine MK 212: agonist 6-chloro-2-(1-piperazinyl)pyrazine HCl MPH: Methylphenidate NAc: Nucleus accumbens NE: Norepinephrine NET: Norepinephrine transporter vii NSD-1015: 3-Hydroxybenzylhydrazine PFC: prefrontal cortex p-MPPF: 4-(2'-methoxy-)-phenyl-1-[2'-(N-2''-pyridinyl)-p-fluorobenzamido-]ethyl- piperazine RU 24969: 5-methoxy-3-(1,2,3,6-tetrahydro-4-pyridinyl)-1H-indole SR 46349B: 1(Z)-[2-(dimethylamino)ethoxyimino]-1(2-fluorophenyl)-3-(4- hydroxyphenyl)-2(E)-propene SERT: serotonin transporter SSRI: selective serotonin reuptake inhibitor Vmax: rate of transporter uptake VTA: ventral tegmental area WAY 100,635: N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]- N-(2- pyridyl)cyclohexanecarboxamide viii LIST OF TABLES AND FIGURES TABLES Page I. The effects of chronic MPH on immediate and long-term neurobiological measures in adolescents………………………………………………….. 12 II. The effects of chronic MPH on immediate and long-term neurobiological measures in adults. ………………………………………………………. 17 III. The effects of chronic MPH on immediate and long-term behavioral measures in adolescents. …………………………………………………. 22 IV. The effects of chronic MPH on immediate and long-term behavioral measures in adults. ……………………………………………………….. 29 V. Effects of chronic MPH on DAT, D2, and SERT levels in the striatum, and on 5-HT1B receptor levels in the ventral midbrain. ………………… 126 FIGURES CHAPTER II Figure 1. Effects of chronic MPH on behavioral and neurochemical responses to cocaine. ……………………………………………………………….. 128 Figure 2. Effects of chronic MPH on behavioral and neurochemical responses to MDMA. ……………………………………………………………….. 131 Figure 3. Effects of chronic MPH on behavioral and neurochemical measures following administration of an SSRI. …………………………………. 134 Figure 4. Effects of chronic MPH on fluoxetine infused into the VTA. ………... 137 ix Figure 5. Effects of chronic MPH on responses to the 5-HT1B agonist RU 141 24969. …………………………………………………………………. CHAPTER III Figure 1. Effects of Chronic MPH on Locomotor Responses to 5-HT receptor agonists. ……………………………………………………………….. 165 Figure 2. Chronic MPH reverses CPP and NAc DA for 5-HT1A agonist 8-OH- DPAT. …………………………………………………………………. 169 Figure 3. Infusion of 8-OH-DPAT into the VTA has differential effects in MPH- treated animals. ………………………………………………………... 172 Figure 4. Chronic MPH reverses CPA and NAc DA for 5-HT1B agonist RU 24969. …………………………………………………………………. 175 Figure 5. Chronic MPH produces sensitization to infusion of 5-HT1B agonist CP93,129 in the VTA. ………………………………………………… 178 Figure 6. Effects of chronic MPH on neurochemical responses to 5-HT1B antagonist GR 55562. …………………………………………………. 181 Figure 7. 5-HT1B antagonists reverse the effects of fluoxetine infusion in MPH- treated animals. ………………………………………………………… 184 CHAPTER IV Figure 1. Effects of chronic MPH on locomotor activity in the open field during habituation and in response to fluoxetine. …………………………… 209 Figure 2. Effects of chronic MPH on Forced Swim Test behavior. ……………... 212 x Figure 3. Effects of chronic MPH on tail suspension test measures. ……………. 215 Figure 4. Effects of chronic