Long-Lasting Memory Impairments after Subchronic Immune Challenge by Daria Tchessalova A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Neuroscience) in the University of Michigan 2019 Doctoral Committee: Assistant Professor Natalie C. Tronson, Chair Professor Anuska Andjekovic-Zochowska Professor Gabriel Corfas Assistant Professor Jacek Debiec Professor Shigeki Iwase Daria Tchessalova [email protected] ORCID iD: 0000-0001-8619-0922 © Daria Tchessalova 2019 Acknowledgements As a graduate student at the University of Michigan, I have had the exceptional privilege of training in a research-intensive, collaborative, supportive academic environment. I would like to sincerely thank my dissertation mentor, Dr. Natalie Tronson for her close mentorship, endless efforts in guiding me to expand my critical thinking and problem solving skills, encouragement in presenting my graduate work at conferences and presentations, and all of her support in my intellectual growth. Dr. Tronson has taught me to exercise the conceptual framework necessary for effective presentations, grant proposals, and manuscripts. I very much appreciate all of her efforts to help me become the scientist and researcher that I am today. I admire her for her effortless ability to efficiently achieve long-term project goals and to assist students with any issues, even last minute issues, related to their research projects or personal lives. I would like to thank the members of my committee for all of their wonderful suggestions regarding my research project throughout the years. Dr. Jacek Debiec has been a very helpful and encouraging mentor. His philosophical approach to science inspires me to think beyond the scope of specific research questions in my project to better understand how my research impacts individuals suffering from various neurological disorders and diseases. I would like to sincerely thank Dr. Gabriel Corfas for allowing me to explore research questions that I was most passionate about in the field of neuron-glia interactions as a rotating student. While I had not ii received the results that I had hoped for, I learned to effectively design experiments, troubleshoot, and, eventually, avoid letting my stubbornness get in the way of my progress. I would also like to thank Dr. Corfas for his extremely helpful feedback on my dissertation project for my 4th year talk. I would like to sincerely thank our collaborators, Dr. Anuska Andjelkovic- Zochowska and Dr. Svetlana Stamatovic for all of their wonderful guidance and their help with the blood-brain barrier studies as well as for all of the interesting discussions regarding aging and Alzheimer’s disease. I have very much enjoyed our collaboration on cognitive dysfunction in aging and the role of Sirt1. I would also like to very much thank Dr. Shigeki Iwase, Christina Vallianatos, and Tricia Garay for their advice, support, and assistance with prepping of the RNA samples. We would very much like to thank them for allowing us to use their laboratory equipment for RNA extraction. I would also like to thank Dr. Iwase for all of his mentorship and advice for organizing the 2018 NGP Spring Symposium on Neuroepigenetics. Drs. Richard McEachin, Rebecca Tagett, and Robert Lyons from the Bioinformatics for at the University of Michigan have been a huge asset to our gene expression studies. They have advised us regarding the best sequencing method for the RNA-sequencing experiments and performed the Bioinformatics analysis for differentially expressed genes. I would also like to thank Dave Bridges for performing the TRANSFAC analysis in R for predicted transcription factors for our differently expressed genes set. Throughout my time at the University of Michigan, I have been very fortunate to get to know some of the nicest and most helpful co-workers I have ever met. I would like to thank Dr. Ashley Keiser for all her help with my research project over the years as well as her great jokes and supportive nature with any challenges that lay ahead. I would like to very much thank Lacie iii Turnbull for her endless assistance, support, and caring nature that very much helped me get oriented when I first started in the lab. Dr. Katie Collette was also one of the most supportive and caring research and writing mentors and life coaches I have met. I would like to thank Caitlin Posillico for all of her helpful guidance and wonderful conversations related to our research projects and beyond as well as her kind introductions to various members of the Psychoneuroimmunology Research Society. While only in the lab for a short time, Melanie Gil has been an amazing emotional and physical support system with a great passion and can do attitude that has helped me to carry on, even during the toughest days. Brynne Raines has been a very helpful and dedicated lab manager. I would like to thank the undergraduate students who have provided excellent assistance with my research project, especially Grayson Buning who performed behavioral scoring of depressive-like behaviors, and imaging, and cell counting for c-Fos analysis, Rachel Fenberg who completed hand-scoring analysis of various behavioral tests, as well as Faith Best who helped me with analysis of behavioral testing, blood-brain barrier permeability, and microglial activation experiments. Additionally, I would like to thank Eugene Shim for his positive attitude and entertaining nature. iv TABLE OF CONTENTS ACKNOWLEDGEMENTS ............................................................................................. ii TABLE OF CONTENTS ..................................................................................................v LIST OF TABLES .......................................................................................................... vii LIST OF FIGURES ....................................................................................................... viii ABSTRACT ...................................................................................................................... xi Chapter I. Introduction .....................................................................................................1 Long-Lasting Consequences of Systemic Inflammatory Event on Memory and Emotion ........................................................................................1 Neuroimmune Activation after Systemic Inflammatory Event and its Consequences on Behavior .........................................................................4 Memory: Types of Memory, Circuits, and Molecular Mechanisms ........................8 Immune Modulation of Memory and Cognitive Functions ..................................10 Mechanisms of memory dysfunction after systemic inflammatory event ............14 Goal Dissertation Project and Summary Hypotheses ............................................17 Chapter II. Long-lasting changes in memory after subchronic immune challenge 20 Abstract ..................................................................................................................20 Introduction ............................................................................................................21 Materials and methods ...........................................................................................23 Results ....................................................................................................................27 Discussion ..............................................................................................................36 Figures....................................................................................................................43 Chapter III. Implications of subchronic immune challenge on long-lasting depressive-like and anxiety-like behaviors ...................................................................53 Abstract ..................................................................................................................53 Introduction ............................................................................................................54 Materials and methods ...........................................................................................56 Results ....................................................................................................................60 Discussion ..............................................................................................................66 Figures....................................................................................................................74 Chapter IV. Enduring and sex-specific changes in hippocampal gene expression after subchronic immune challenge ...............................................................................84 Abstract ..................................................................................................................84 Introduction ............................................................................................................85 Materials and methods ...........................................................................................87 Results ....................................................................................................................91 Discussion ............................................................................................................104 Tables ...................................................................................................................113