University of Pennsylvania ScholarlyCommons Publicly Accessible Penn Dissertations 2016 Sex, Drugs, and Transgenerational Inheritance: Are the Kids Alright? Nicole Lynn Yohn University of Pennsylvania, [email protected] Follow this and additional works at: https://repository.upenn.edu/edissertations Part of the Neuroscience and Neurobiology Commons Recommended Citation Yohn, Nicole Lynn, "Sex, Drugs, and Transgenerational Inheritance: Are the Kids Alright?" (2016). Publicly Accessible Penn Dissertations. 2118. https://repository.upenn.edu/edissertations/2118 This paper is posted at ScholarlyCommons. https://repository.upenn.edu/edissertations/2118 For more information, please contact [email protected]. Sex, Drugs, and Transgenerational Inheritance: Are the Kids Alright? Abstract The occurrence of neuropsychiatric disorders and substance abuse is subject to familial inheritance (nature) as well influence from the environment (nurture). In addition, familial patterns of behavior and disease are also mediated by transgenerational epigenetic inheritance. The dynamic nature of the epigenome allows for exposures to stress, drugs of abuse, environmental toxins, and even changes in diet to produce changes in gene expression. In addition these changes can be inherited by offspring. Therefore, offspring behavior and quality of life are shaped by their own experiences as well as the experiences of their parents and more distant relatives. The studies in this dissertation had two objectives: first ot identify the transgenerational inheritance of adolescent stress exposure and its effects on offspring behavior including response to nicotine and second, to determine the cross-generational interaction of nicotine and stress exposures on offspring behavior. To address the first objective, we exposed male and female mice to adolescent stress exposure, determined the long-term effects of exposure on their behavior, and identified changes in phenotype in their F1 and F2 offspring. eW found that adolescent stress exposure produced changes in anxiety, startle response, and gene expression in adulthood that was not found when the same stress exposure occurred in adult mice. In addition, we found that transgenerational inheritance of adolescent stress exposure promoted sex- and lineage- dependent changes in anxiety, depression, startle, and response to nicotine in F1 and F2 offspring. Furthermore, to determine if parental stress exposure influenced gene expression in the brains of offspring we analyzed the transcriptome of F1 males and found 240 differentially expressed genes in the amygdala of males whose fathers were exposed to stress. In our final study, we developed a novel multigenerational exposure paradigm and determined that F0 nicotine and F1 stress exposure interact across generations to produce unique phenotypes in F2 and F3 offspring. Together, research from this dissertation provides evidence of an adolescent chronic stress exposure that mediates anxiety in adulthood and is inherited in future generations by reprogramming the brain of offspring, and provides the first example of cross-generational interactions of two environmental exposures to influence offspring phenotype. Degree Type Dissertation Degree Name Doctor of Philosophy (PhD) Graduate Group Neuroscience First Advisor Julie A. Blendy Keywords Behavior, Chronic stress, Drug abuse, Inheritance, Mouse, Nicotine Subject Categories Neuroscience and Neurobiology This dissertation is available at ScholarlyCommons: https://repository.upenn.edu/edissertations/2118 SEX, DRUGS, AND TRANSGENERATIONAL INHERITANCE: ARE THE KIDS ALRIGHT? Nicole Lynn Yohn A DISSERTATION in Neuroscience Presented to the Faculties of the University of Pennsylvania in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy 2016 Supervisor of Dissertation Graduate Group Chairperson ________________________ _______________________ Julie A. Blendy, PhD Joshua I. Gold, PhD Professor of Pharmacology Professor of Neuroscience Dissertation Committee Jon Lindstrom, PhD, Professor of Neuroscience Caryn Lerman, PhD, Professor of Psychiatry Irwin Lucki, PhD, Professor of Pharmacology Marsia S. Bartolomei, PhD, Professor of Cell and Developmental Biology i To the women in science who inspired me to pursue my degree: Martha Kermizis, Dr. Lina Yoo, Dr. Susan Kennedy, and Dr. Julie Blendy. And to the most inspiring woman I know, Lisa Yohn. ii ACKNOWLEDGMENT First, I would like to express my sincere gratitude to my advisor Dr. Julie Blendy for the continuous support of my PhD study and related research, for her patience, motivation, and immense knowledge. Her guidance helped me in all the time of research and writing of this thesis, and in many moments her selfless time and care were sometimes all that kept me going. I could not have imagined having a better advisor and mentor for my PhD study. Besides my advisor I would like to thank the rest of my thesis committee: Dr. Jon Lindstrom, Dr. Caryn Lerman, Dr. Irwin Lucki, and Dr. Marisa Bartolomei, for their insightful comments and encouragement, but also for the hard questions which incented me to widen my rsearch from various perspectives. My sincere thanks also goes to Dr. Klaus Keastner, Dr. Diana Bernstein, the Bartolemei Lab, and the Next Generation Sequencing Core who provided me with collaboration and access to their research knowledge and facilities. In addition, I would like to thank Dr. Yemin Lan and Dr. Jonathan Schug for their expertise in data analysis. Without there precious support it would not have been possible to conduct this research. I thank my fellow labmates for the stimulating discussions, the countless hours spent in the animal colony working together, and for all the fun we had over the past five years. In particular, I am grateful for Dr. Bridgin Lee, without whom this would not have been possible. I am lucky to be leaving graduate school with one of the best friends I will have made in my entire life. Not only did she challenge me to be a better scientist but also taught me poise, grace, and courage in the process. I would like to thank my fellow iii “Penn Pals” in the Neuroscience Graduate Group and BGS for their support, shoulders to lean on, and countless memories made together. Finally thank you to Liz, Jessie, Britt, and Laura – behind every successful woman is a tribe of women who have her back. Last but not leat, I would like to thank my family: my parents and husband Jason. There unwavering support has propelled me to places I never thought possible. Thank you Jason for reading every page of this thesis and being with me every step of the way! To my extended family, thank you for supporting me each and every day, before, during, and after graduate school. iv ABSTRACT SEX, DRUGS, AND TRANSGENERATIONAL INHERITANCE: ARE THE KIDS ALRIGHT? Nicole L. Yohn Julie A. Blendy The occurrence of neuropsychiatric disorders and substance abuse is subject to familial inheritance (nature) as well influence from the environment (nurture). In addition, familial patterns of behavior and disease are also mediated by transgenerational epigenetic inheritance. The dynamic nature of the epigenome allows for exposures to stress, drugs of abuse, environmental toxins, and even changes in diet to produce changes in gene expression. In addition these changes can be inherited by offspring. Therefore, offspring behavior and quality of life are shaped by their own experiences as well as the experiences of their parents and more distant relatives. The studies in this dissertation had two objectives: first to identify the transgenerational inheritance of adolescent stress exposure and its effects on offspring behavior including response to nicotine and second, to determine the cross-generational interaction of nicotine and stress exposures on offspring behavior. To address the first objective, we exposed male and female mice to adolescent stress exposure, determined the long-term effects of exposure on their behavior, and identified changes in phenotype in their F1 and F2 offspring. We found that adolescent stress exposure produced changes in anxiety, startle response, and gene expression in adulthood that was not found when the same stress exposure occurred in v adult mice. In addition, we found that transgenerational inheritance of adolescent stress exposure promoted sex- and lineage-dependent changes in anxiety, depression, startle, and response to nicotine in F1 and F2 offspring. Furthermore, to determine if parental stress exposure influenced gene expression in the brains of offspring we analyzed the transcriptome of F1 males and found 240 differentially expressed genes in the amygdala of males whose fathers were exposed to stress. In our final study, we developed a novel multigenerational exposure paradigm and determined that F0 nicotine and F1 stress exposure interact across generations to produce unique phenotypes in F2 and F3 offspring. Together, research from this dissertation provides evidence of an adolescent chronic stress exposure that mediates anxiety in adulthood and is inherited in future generations by reprogramming the brain of offspring, and provides the first example of cross-generational interactions of two environmental exposures to influence offspring phenotype. vi TABLE OF CONTENTS ACKNOWLEDGMENT ............................................................................................................. III ABSTRACT .................................................................................................................................