Loyola University Chicago Loyola eCommons Dissertations Theses and Dissertations 2018 Mechanisms and Consequences of Epigenetic Inheritance Following Parental Preconception Alcohol Exposure Annadorothea Asimes Loyola University Chicago Follow this and additional works at: https://ecommons.luc.edu/luc_diss Part of the Neurosciences Commons Recommended Citation Asimes, Annadorothea, "Mechanisms and Consequences of Epigenetic Inheritance Following Parental Preconception Alcohol Exposure" (2018). Dissertations. 2770. https://ecommons.luc.edu/luc_diss/2770 This Dissertation is brought to you for free and open access by the Theses and Dissertations at Loyola eCommons. It has been accepted for inclusion in Dissertations by an authorized administrator of Loyola eCommons. For more information, please contact [email protected]. Copyright © 2018 Annadorothea Asimes LOYOLA UNIVERSITY CHICAGO MECHANISMS AND CONSEQUENCES OF EPIGENETIC INHERITANCE FOLLOWING PARENTAL PRECONCEPTION ALCOHOL EXPOSURE A DISSERTATION SUBMITTED TO THE FACULTY OF THE GRADUATE SCHOOL IN CANDIDACY FOR THE DEGREE OF DOCTOR OF PHILOSOPHY PROGRAM IN NEUROSCIENCE BY ANNADOROTHEA ASIMES CHICAGO, IL MAY 2018 Copyright by AnnaDorothea Asimes, 2018 ALL rights reserved. ACKNOWLEDGEMENTS The work required to compLete this dissertation wouLd not have been possibLe alone. I wouLd first Like to thank my advisor and mentor, Dr. Toni Pak. The guidance and support you have given me throughout these graduate schooL years has been invaluabLe. You have provided a chalLenging environment that pushed me to be resourcefuL, creative, boLd, and humbLed. I wouLd not be the scientist I am today without your mentorship and friendship. I wouLd also Like to thank my Lab mates, who quickLy turned from coworkers to friends. I Look forward to buiLding our Lab famiLy and seeing the great work you alL go on to do. SecondLy, I wouLd Like to thank my dissertation committee for the time and effort you have contributed to my work and personal deveLopment. Each of you have improved this dissertation through your questions and concerns, and I hope to, one day, be abLe to provide this same rigor for future graduate students. FinalLy, I wouLd Like to thank my famiLy for the encouragement to, first and foremost, pursue my education. I have always feLt extremeLy Lucky to have such great and supportive parents, who instiLLed in me a drive to chase my interests, and alLowed me to try even my craziest of ideas. I am now abLe to add my husband to this List of cheerLeaders. You have shown great patience as I worked Long, and often frustrating, hours to realize this dream. My hope is that alL of you enjoy reading this dissertation as much as I enjoyed writing it. I can never thank you alL enough. iii TABLE OF CONTENTS ACKNOWLEDGEMENTS ................................................................................................................. iii LIST OF TABLES ............................................................................................................................. vii LIST OF FIGURES .......................................................................................................................... viii LIST OF ABBREVIATIONS ................................................................................................................. x ABSTRACT ..................................................................................................................................... xii CHAPTER I: STATEMENT OF THE PROBLEM .................................................................................... 1 CHAPTER II: INTRODUCTION .......................................................................................................... 3 Literature Review ....................................................................................................................... 3 Epigenetics: Environment Meets Genetics ............................................................................. 3 DNA MethyLation – Writers, Readers, and Erasers. ................................................................ 4 Functional Meaning of DNA in Different Parts of the Genome. ............................................. 7 Age-Driven Changes in Somatic MethyLation. ......................................................................... 9 Epigenetic Reprogramming During Embryonic DeveLopment. ............................................. 10 MuLtigenerational Epigenetic Inheritance. ........................................................................... 12 AdoLescence: DeveLopment and Drinking. ............................................................................ 15 Inheritance of Parental ALcohoL Exposure. ............................................................................ 17 MoLecuLar Actions of Alcohol in Somatic CeLLs- Non-Genomic. ............................................. 18 MoLecuLar Actions of Alcohol in Somatic CeLLs– Genomic and Epigenomic. ......................... 21 AlcohoL and the Hypothalamus. ............................................................................................ 25 Summary ................................................................................................................................... 28 CHAPTER III: ADOLESCENT BINGE-PATTERN ALCOHOL EXPOSURE ALTERS GENOME WIDE DNA METHYLATION PATTERNS IN THE HYPOTHALAMUS OF ALCOHOL NAÏVE MALE OFFSPRING ...... 32 Introduction .............................................................................................................................. 32 ResuLts ...................................................................................................................................... 35 Parental EthanoL Exposure Induces DifferentialLy MethyLated Cytosine Residues in the Hypothalamus Of Male EthanoL-Naïve Offspring. ................................................................. 35 DifferentialLy MethyLated Cytosines Were Distributed Across ALL Chromosomes and the Extent of Hypo- Versus HypermethyLation was Dependent on Parental EthanoL Exposure. 41 DMCs were PrimariLy Observed Outside Of CpG IsLands and in Intergenic Regions. ............ 46 mRNA Expression of DifferentialLy MethyLated Genes is ALtered in the Hypothalamus. ...... 49 Discussion ................................................................................................................................. 52 CHAPTER IV: PARENTAL PRECONCEPTION ALCOHOL EXPOSURE IMPACTS OFFSPRING DEVELOPMENT THROUGH PUBERTY ............................................................................................ 59 Introduction .............................................................................................................................. 59 iv ResuLts ...................................................................................................................................... 61 Experiment 1: BaseLine Characteristics of Male and Female Offspring from Parents Administered Preconception Binge-Pattern EtOH During Adolescence (Fig 9-13). .............. 61 Experiment 2: Effects of AdoLescent EtOH Exposure in Offspring with Parental History of Preconception EtOH Consumption (Fig 14-18). .................................................................... 69 Experiment 3: Tracking Transcriptional ReguLation of Candidate Genes in the Offspring Hypothalamus Through DeveLopment FoLLowing Parental EtOH Exposure (TabLes 7, 8). ..... 77 Discussion ................................................................................................................................. 80 CHAPTER V: "FATTY BRAIN" FOLLOWING PUBERTAL BINGE ETHANOL EXPOSURE ..................... 88 Introduction .............................................................................................................................. 88 ResuLts ...................................................................................................................................... 91 ApoE was Increased in AduLt Males FoLLowing Pubertal EtOH Exposure. ............................. 91 Lipid-Associated Genes are Unaltered in JuveniLe and AdoLescent Offspring Hypothalamus. .............................................................................................................................................. 92 Primary Astrocytes CuLtured from AduLt Males Showed Increased ChoLesteroL AccumuLation. ....................................................................................................................... 93 Fatty Astrocytes are Less Neuroprotective Against Toxic InsuLt. .......................................... 96 Discussion ................................................................................................................................. 97 CHAPTER VI: FINAL DISCUSSION ................................................................................................. 101 Summary ................................................................................................................................. 101 Key Findings ............................................................................................................................ 103 Final Thoughts ........................................................................................................................ 106 Maternal AND Paternal Preconception Behaviors InfLuence Offspring. ............................. 106 Epigenomics is InvaluabLe, but Cannot Give the WhoLe Picture Yet. .................................. 107 Epigenetic Inheritance is not