The Effect of Dietary Fat on Obesity, Gene Expression, and DNA Methylation in Two Generations of Mice" (2017)
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Washington University in St. Louis Washington University Open Scholarship Arts & Sciences Electronic Theses and Dissertations Arts & Sciences Summer 8-15-2017 The ffecE t of Dietary Fat on Obesity, Gene Expression, and DNA Methylation in Two Generations of Mice Madeline Rose Keleher Washington University in St. Louis Follow this and additional works at: https://openscholarship.wustl.edu/art_sci_etds Part of the Human and Clinical Nutrition Commons, and the Physiology Commons Recommended Citation Keleher, Madeline Rose, "The Effect of Dietary Fat on Obesity, Gene Expression, and DNA Methylation in Two Generations of Mice" (2017). Arts & Sciences Electronic Theses and Dissertations. 1219. https://openscholarship.wustl.edu/art_sci_etds/1219 This Dissertation is brought to you for free and open access by the Arts & Sciences at Washington University Open Scholarship. It has been accepted for inclusion in Arts & Sciences Electronic Theses and Dissertations by an authorized administrator of Washington University Open Scholarship. For more information, please contact [email protected]. WASHINGTON UNIVERSITY IN ST. LOUIS Division of Biology and Biomedical Sciences Evolution, Ecology, and Population Biology Dissertation Examination Committee: Jim Cheverud, Chair Jenny Duncan Allan Larson Kelle Moley Ken Olsen Ting Wang The Effect of Dietary Fat on Obesity, Gene Expression, and DNA Methylation in Two Generations of Mice by Madeline Rose Keleher A dissertation presented to The Graduate School of Washington University in partial fulfillment of the requirements for the degree of Doctor of Philosophy August 2017 St. Louis, Missouri © Copyright 2017 by Madeline Rose Keleher. All rights reserved. TABLE OF CONTENTS List of Tables…………………………………………………………………………………….iv List of Figures…………………………………………………………………………………...vi Acknowledgements…………………………………………………………………………….viii Abstract of the Dissertation…………………………………………………………………...xii Chapter 1. Introduction of the Dissertation…………………………………………………....1 Chapter 2. The Effect of Dietary Fat on Behavior in Mice…………………………………..19 Abstract…………………………………………………………………………………..20 Introduction………………………………………………………………………………20 Methods…………………………………………………………………………….…….25 Results……………………………………………………………………………………29 Discussion………………………………………………………………………….…….32 Literature Cited……………………………………………………………….………….36 Tables and Figures……………………………………………………….………………41 Chapter 3. A High-Fat Diet Alters Genome-wide DNA Methylation and Gene Expression in SM/J Mice………………………………………………………………...48 Abstract………………………………………………………………………………..…49 Introduction………………………………………………………………………………49 Methods…………………………………………………………………………….…….54 Results……………………………………………………………………………………59 Discussion………………………………………………………………………….…….63 Literature Cited……………………………………………………………….………….71 Tables and Figures……………………………………………………….………………79 ii Chapter 4. Maternal Obesity Alters Offspring Gene Expression, DNA Methylation, and Obesity Risk in Mice……...……………………………………….....97 Abstract…………………………………………………………………………………..98 Introduction………………………………………………………………………………98 Methods…………………………………………………………………………….…...104 Results…………………………………………………………………………………..111 Discussion………………………………………………………………………….…...117 Literature Cited……………………………………………………………….………...129 Tables and Figures……………………………………………………….……………..142 Chapter 5. Conclusion of the Dissertation…………………………………………………...186 iii LIST OF TABLES Table 2.1……………………………………………………………………………………..…..42 Table 2.2……………………………………………………………………………………..…..43 Table 2.3……………………………………………………………………………………..…..44 Table 3.1……………………………………………………………………………………..…..79 Table 3.2……………………………………………………………………………………..…..80 Table 3.3……………………………………………………………………………………..…..84 Table 3.4……………………………………………………………………………………..…..84 Table 3.5……………………………………………………………………………………..…..84 Table 3.6……………………………………………………………………………………..…..85 Table 3.7……………………………………………………………………………………..…..85 Table 3.8……………………………………………………………………………………..…..86 Table 3.9……………………………………………………………………………………..…..87 Table 3.10……………………………………………………………………………………..…91 Table 3.11……………………………………………………………………………………..…96 Table 4.1……………………………………………………………………………………..…142 Table 4.2……………………………………………………………………………………..…143 Table 4.3……………………………………………………………………………………..…144 Table 4.4……………………………………………………………………………………..…144 Table 4.5……………………………………………………………………………………..…148 Table 4.6……………………………………………………………………………………..…150 Table 4.7……………………………………………………………………………………..…157 Table 4.8……………………………………………………………………………………..…158 iv Table 4.9………………………………………………………………………………………..162 Table 4.10…………………………………………………………………………………..…..164 Table 4.11………………………………………………………………………………..……..165 Table 4.12…………………………………………………………………………………..…..165 Table 4.13………………………………………………………………………………..……..168 Table 4.14………………………………………………………………………………..……..168 Table 4.15………………………………………………………………………………..……..170 Table 4.16………………………………………………………………………………..……..172 Table 4.17………………………………………………………………………………..……..174 Table 4.18……………………………………………………………………..………………..175 v LIST OF FIGURES Figure 2.1………………………………………………………………………………………...43 Figure 2.2 ………………………………………………………………………………………..43 Figure 2.3…………………………………………………………………………………….…..45 Figure 2.4………………………………………………………………………………………...45 Figure 2.5………………………………………………………………………………………...46 Figure 2.6………………………………………………………………………………………...47 Figure 2.7………………………………………………………………………………………...47 Figure 3.1………………………………………………………………………………………...81 Figure 3.2………………………………………………………………………………………...82 Figure 3.3………………………………………………………………………………………...83 Figure 3.4………………………………………………………………………………………...84 Figure 3.5………………………………………………………………………………………...88 Figure 3.6………………………………………………………………………………………...89 Figure 3.7………………………………………………………………………………………...90 Figure 3.8………………………………………………………………………………………...95 Figure 4.1……………………………………………………………………………………….142 Figure 4.2……………………………………………………………………………………….142 Figure 4.3……………………………………………………………………………………….145 Figure 4.4……………………………………………………………………………………….146 Figure 4.5……………………………………………………………………………………….147 Figure 4.6……………………………………………………………………………………….149 Figure 4.7……………………………………………………………………………………….159 vi Figure 4.8……………………………………………………………………………………….160 Figure 4.9……………………………………………………………………………………….161 Figure 4.10……………………………………………………………………………………...164 Figure 4.11……………………………………………………………………………………...166 Figure 4.12……………………………………………………………………………………...167 vii ACKNOWLEDGEMENTS First and foremost, I would like to thank Jim Cheverud for being my advisor and supporting me these six long years. Jim, your comments on my earlier drafts of this dissertation made the final product far better. I am also indebted to my committee members. I had never done wet lab work before my lab rotation with Jenny Duncan, my co-advisor who personally showed me how to use a micropipettor and extract DNA, and patiently taught me how to troubleshoot when I made mistakes. Allan Larson has given me wise advice and is one of the best teachers I’ve ever had. Kelle Moley gave me good feedback and helped when I was having difficulties getting the high-fat-fed mice to reproduce. Ting Wang is charismatic and has endless energy for science; he introduced me to epigenetics and has been a great role model for how to give an engaging science talk. Ken Olsen always managed to respond to my emails within 10 minutes, submitted many letters of recommendation for me, helped me feel connected to Wash U’s program even during the four years I spent in Chicago, and is a superb teacher and scientist. A special thank you to my middle school science teacher, Jim Gravagna, who nurtured my interest in science and convinced me to join the school Science Olympiad team. He is a gifted teacher and an amazing coach. Thank you to Charles Roseman, who started my career in research by letting me into his lab when I was a sophomore at the University of Illinois. What I thought was just a semester-long honors project turned into a 2.5-year research position with a hilarious, brilliant scientist. It hadn’t occurred to me to do research or to apply to graduate school until Charles encouraged me to. I also want to thank Stan Braude, who treated me as a co-teacher instead of just a TA in his human biology class, let me co-author a book with him to teach the general public to be more informed biomedical consumers, inspired me with his creative teaching techniques, and has viii given me so much support and encouragement over the years. I am extremely thankful for the members of the Cheverud lab in both its St. Louis and Chicago incarnations, including Eric Schmidt, Bing Wang, and Susan Pletscher (Susan was the first person who got to know me in St. Louis and she taught me how to take care of mice). Mihaela Pavlicev has been a mentor and a friend, and I am grateful for her guidance and her eternally cheerful attitude. I also want to thank my wonderful team of undergrads, who worked tirelessly to help collect data for my thesis: Lauren Hicks, Rabab Zaidi, Amer Ahmed, Cassondra Pavlatos, Kayna Patel, Shyam Shah, Elsa Oakley, Carlee Bettler, and Samir El Idrissi. Lauren and Rabab, on the days when I really didn’t feel up to spending hours glucose testing or necropsying mice, your impressive work ethic and sunny dispositions made the time fly by. I would also like to thank the employees of Loyola’s Animal Care Facility, particularly James Robinson and Beth McGeehan, for ensuring that my mice were well taken care of. Thank you to Chris Calderaro and Virginia Lorenzo for helping me with ordering all the supplies necessary for my research, especially when it was an urgent last-minute request or involved exceedingly complicated paperwork. I am grateful to Catherine Putonti