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(Hh/Gli) Regulates Neurogenesis in the Zebrafish Hypothalamus

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(Hh/Gli) Regulates Neurogenesis in the Zebrafish Hypothalamus University of Massachusetts Amherst ScholarWorks@UMass Amherst Doctoral Dissertations Dissertations and Theses July 2020 Hedgehog Signaling (Hh/Gli) Regulates Neurogenesis in the Zebrafish Hypothalamus Ira Male University of Massachusetts Amherst Follow this and additional works at: https://scholarworks.umass.edu/dissertations_2 Part of the Other Cell and Developmental Biology Commons Recommended Citation Male, Ira, "Hedgehog Signaling (Hh/Gli) Regulates Neurogenesis in the Zebrafish Hypothalamus" (2020). Doctoral Dissertations. 1952. https://doi.org/10.7275/17586512 https://scholarworks.umass.edu/dissertations_2/1952 This Open Access Dissertation is brought to you for free and open access by the Dissertations and Theses at ScholarWorks@UMass Amherst. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of ScholarWorks@UMass Amherst. For more information, please contact [email protected]. HEDGEHOG SIGNALING (Hh/GLI) REGULATES NEUROGENESIS IN THE ZEBRAFISH HYPOTHALAMUS A Dissertation Presented by IRA MALE Submitted to the Graduate School of the University of Massachusetts Amherst in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY May 2020 Molecular and Cellular Biology © Copyright by Ira Male 2020 All Rights Reserved HEDGEHOG SIGNALING (Hh/GLI) REGULATES NEUROGENSIS IN THE ZEBRAFISH HYPOTHALAMUS A Dissertation Presented by IRA MALE Approved as to style and content by: ____________________________________ Rolf O. Karlstrom, Chair ____________________________________ Lawrence M. Schwartz, Member ____________________________________ Alicia R. Timme-Laragy, Member ____________________________________ R. Craig Albertson, Member ____________________________________ Scott C. Garman, Director Program in Molecular and Cellular Biology DEDICATION For my parents, Leftoni and Nesim Male, who knew how to be courageous and humble in their journey to the US. Temen ia dedikoj prinderve te mi, Leftoni dhe Nesim Male. Dedikimi ndaj familjes ne vite eshte frymezim per mua ACKNOWLEDGMENTS Graduate school has been a captivating experience. Few people succeed solely on their own, and I am no exception. There are several people that played a part in my graduate school journey, and I would like to thank many of them here. My thesis advisor, Professor Rolf Karlstrom, invited me to join his group and work on a project that was challenging and insightful. I am appreciative for the guidance and direction through the years. The other members of the Karlstrom lab served as a great group of friends and colleagues throughout my time there. I am thankful for the willingness and availability of Dr. Mary McKitrick to teach me everything she knew and letting me borrow her beautifully written lab notebooks. She is a mentor and a true friend. Two undergraduate students directly assisted with my project. Rubi Tanimura helped with the TET lines and performed many immunofluorescence experiments with me. Matt Loring helped with the quantification of different experiments and was always available for anything computer related tasks. These last two years, Grace Lawson, has graciously helped with lab tasks such as fish sorting and solution making, freeing up a lot of time that I used thinking about my science. I am also thankful to Anna Aristarkhova for her help in the lab this summer. I have truly enjoyed Dr. Priya Gosh comradery since she joined the lab and I appreciate her willingness to prioritize collaboration over competition. My many friends at UMass have made life outside the lab a joy. Alyssa and I shared a love for late night lab work and Italian gelato. Trivia nights with Trish where our love for random facts won us ramen noodles by placing dead last have produced V a lot of laughter through the years. The trip Liv and I took to Pennsylvania to get five gallons of breast milk for her research was epic – great memories. To Lisa, I am grateful for all the help through the years. My dissertation advisory committee members, Prof. Larry Schwartz, Prof. Alicia Timme- Laragy, and Prof. Craig Albertson were always willing to meet and I am thankful for their valuable input. The Molecular and Cellular Program provided a supportive environment for my graduate studies. To Carrie, Sarah and Jimmy I am thankful for their help with teaching assistant assignment scheduling, funding paperwork, and so much more. I would also like to thank several instructors in the Biology department for whom I was a teaching assistant during my five years at UMass; Prof. Laura Francis, Prof. Elizabeth Connor, Prof. Kari Loomis were all great to work for. Dr. Kate Dorfman was invaluable from helping my teaching set-ups for different classes to stepping in my place when my father was in the hospital. I am also thankful to the office of professional development for their help through the years. Outside of UMass, many others deserve to be recognized. To Dr. Bryan, my first scientific advisor in college, I am thankful for his motivation and guidance in learning anything angiogenesis related. To Dr. Pederson, I am forever thankful for him being only an e-mail away and providing his insightful mentorship, direction, and help whenever I needed it. The Marine Biological Laboratories provided a very supportive scientific environment during my attendance in the 2016 Embryology class. I am thankful to the heavily invested course directors, Dr. Behringer and Dr. Alvarado-Sanchez and to the amazing course instructors that made the 6-weeks in Woods Hole scientifically magical. Embryo2016 are friends for life. vi Finally, I would like to thank my family. My sister, has been there for me from the beginning; when she first gifted me her prized dictionary so I could translate and do my high school English homework to the continuous present encouragement. She has graced our family with two additional angels, my nephews Adrian and Albin that are the joy and pride of my parents or the “joint-chiefs” of mischief for me. I am thankful to her and my brother-in-law for all the fun and adventures they have provided me with every time I have visited them in Sweden. My parents embarked on a tough journey to come to the United States, in their 50s with no prior knowledge of the English language or culture, for a better future for my sister and I. Placing me in educational environments where my talents had the opportunity to grow, and the selfless and unconditional encouragement they have provided through the years, are responsible for anything and everything I have accomplished. We immigrated to the States by winning the green card lottery but I consider being born in this family winning the lottery of a lifetime. vii ABSTRACT HEDGEHOG SIGNALING (Hh/GLI) REGULATES ZEBRAFISH HYPOTHALAMIC PROLIFERATION MAY 2020 IRA MALE, B.S., WORCESTER STATE COLLEGE Ph.D., UNIVERSITY OF MASSACHUSETTS AMHERST Directed by: Professor Rolf O. Karlstrom The hypothalamus is the central regulator of a diverse homeostatic processes that are essential for the survival of the animal. Recently, the hypothalamus has been shown to be a neurogenic zone in many animal species including mice, zebrafish, sheep and potentially even humans with hypothalamic neurogenesis implicated in energy metabolism, stress, anxiety and reproduction. However, the identification of the cellular components and the signaling mechanisms regulating hypothalamic neurogenesis are only beginning to be elucidated. My dissertation research elucidates the role of Hh signaling in regulating hypothalamic neurogenesis throughout life. The first chapter of the dissertation offers a survey into the field of neurogenesis with a focus on up-to-date information about hypothalamic neurogenesis. Chapter two demonstrates the role of Hh signaling in the ventral forebrain proliferation. More specifically, we reveal for the first time, that Hh signaling is necessary and sufficient for regulating hypothalamic proliferation rates throughout life. Additionally, we show Hh-responsive radial glia as multipotent neural progenitors giving rise to dopaminergic, GABA-ergic and serotonergic neuronal lineages with Hh signaling being both necessary and sufficient for regulating larval serotonergic cell numbers. Chapter three of my dissertation focuses on a detailed description of the proliferative viii rates of five progenitor cell types present at the hypothalamic lateral (LR) and posterior (PR) recesses. We show that approximately, 60% of Hh-responsive cells in the PR were PCNA positive compared to approximately 20% of Notch-responsive cells and 10% of Shh-producing cells and no proliferative Wnt-responsive. These analyses reveal striking differences in the proliferative profiles among radial glia in the posterior recess with Hh-responsive cells representing a much more proliferative population that may account for the majority of hypothalamic growth in the larval stages. Lastly, chapter four of the dissertation offers a synthesis of the hypothalamic neurogenesis with a focus on the heterogeneity of the neural progenitor types on this brain region. Taken together, my dissertation work reveals a novel role for Hh signaling as a key molecular regulator of hypothalamic proliferation in the zebrafish hypothalamus and provides a foundation for future studies of a mechanism of action. ix TABLE OF CONTENTS Page ACKNOWLEDGMENTS ...................................................................................................5 ABSTRACT ....................................................................................................................
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