Understanding SCN8A and Voltage- Gated Sodium Channelopathies Item Type text; Electronic Dissertation Authors Encinas, Alejandra Carmen Publisher The University of Arizona. Rights Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction, presentation (such as public display or performance) of protected items is prohibited except with permission of the author. Download date 05/10/2021 10:12:59 Link to Item http://hdl.handle.net/10150/634330 1 UNDERSTANDING SCN8A AND VOLTAGE-GATED SODIUM CHANNELOPATHIES By Alejandra del Carmen Encinas Copyright © Alejandra del Carmen Encinas 2019 A Dissertation Submitted to the Faculty of the GRADUATE INTERDISCIPLINARY PROGRAM IN GENETICS In Partial Fulfillment of the Requirements For the Degree of DOCTOR OF PHILOSOPHY In the Graduate College THE UNIVERSITY OF ARIZONA 2019 2 THE UNIVERSITY OF ARIZONA GRADUATE COLLEGE As members of the Dissertation Committee, we certify that we have read the dissertation prepared by Alejandra del Carmen Encinas, titled Understanding SCN8A and Voltage-Gated Sodium Channelopathies and recommend that it be accepted as fulfilling the dissertation requirement for the Degree of Doctor of Philosophy. _________________________________________________________________ Date: August 13, 2019 Michael Hammer _________________________________________________________________ Date: August 13, 2019 Thomas Doetschman _________________________________________________________________ Date: August 13, 2019 Ida (Ki) Moore _________________________________________________________________ Date: August 13, 2019 Todd Vanderah _________________________________________________________________ Date: August 13, 2019 Joseph Watkins Final approval and acceptance of this dissertation is contingent upon the candidate’s submission of the final copies of the dissertation to the Graduate College. I hereby certify that I have read this dissertation prepared under my direction and recommend that it be accepted as fulfilling the dissertation requirement. _________________________________________________________________ Date: August 13, 2019 Michael Hammer Dissertation Committee Chair Department of Genetics 3 STATEMENT BY AUTHOR This dissertation has been submitted in partial fulfillment of requirements for an advanced degree at the University of Arizona and is deposited in the University Library to be made available to borrowers under rules of the Library. Brief quotations from this dissertation are allowable without special permission, provided that an accurate acknowledgement of the source is made. Requests for permission for extended quotation from or reproduction of this manuscript in whole or in part may be granted by the head of the major department of the Dean of the Graduate College when in his or her judgment the proposed use of the material is in the interests of scholarship. In all other instances, however, permission must be obtained from the author. SIGNED: Alejandra del Carmen Encinas 4 ACKNOWLEDGEMENTS There are many people who supported me during my time in the Genetics Graduate Interdisciplinary Program. First of all, I would like to thank my advisor, Dr. Michael Hammer, who introduced me to such an amazing and welcoming patient community that made my research and time in his lab so incredibly meaningful. To Dr. Joseph Watkins, who I met years ago as an undergrad, thank you for guiding me through my research program and always being available to answer a question or five that would pop into my head. To Ki, who I always found to be a rock for me when I was feeling discouraged. To Dr. Thomas Doetschman, whose knowledge and expertise was invaluable as we tried to navigate the mysteries of our mouse colony. And last by certainly not least, to Dr. Todd Vanderah, your insights and the welcoming nature of your own lab made research so fun, even when I was creating 100 miniature peanut butter pellets. I would also like to thank members working within labs that I collaborated with during my time here. To Tally, who always made herself available for my questions or to show me new procedures, thank you for looking out for me. To Tama, who cared for our mouse colony, thank you for all the time you spent answering my questions or giving me suggestions and for your willingness to help out in any way possible. To Connie, thank you for your truth and for never holding back your opinion, I strive to be as confident as you one day. Next, I would like to thank Xenon Pharmaceuticals and the National Institutes of Health (NIH) for their financial support. Xenon provided me with such an amazing opportunity to focus on my research at the very start of my time in Dr. Hammer’s lab. Their feedback and passion on my research was an experience I will never forget. For my NIH grant I would like to thank Dr. 5 Timothy Secomb, who was a fantastic teacher and advisor that taught me new ways to approach modeling in biological systems. Additionally, I could not have completed this program without the support from my family and friends. My mom, Rosi, for constantly rooting me on and giving me so much confidence. My dad, Carlos, for being proud of me no matter what and always eager to hear about my research. My sister, Adriana, for always celebrating the little accomplishments with me and constantly cheering me on. My husband, Jamison, for always being there for me and supporting me. And finally, I want to thank my best friend, Kelsey, for her constant care packages and pick me ups. 6 DEDICATION To all the families of children with an SCN8A mutation, without their tremendous support and passion for research, none of this would have been possible. 7 TABLE OF CONTENTS ABSTRACT .................................................................................................................................................. 9 INTRODUCTION ...................................................................................................................................... 11 Figures .................................................................................................................................................... 24 Tables ...................................................................................................................................................... 27 References ............................................................................................................................................... 42 PRESENT STUDY ..................................................................................................................................... 44 FUTURE WORK ........................................................................................................................................ 47 APPENDIX A ............................................................................................................................................. 50 Introduction ............................................................................................................................................. 54 Methods .................................................................................................................................................. 56 Results ..................................................................................................................................................... 58 Discussion ............................................................................................................................................... 64 References ............................................................................................................................................... 73 Figures .................................................................................................................................................... 77 Tables ...................................................................................................................................................... 81 Supplementary Material .......................................................................................................................... 83 APPENDIX B ........................................................................................................................................... 100 Introduction ........................................................................................................................................... 105 Methods ................................................................................................................................................ 106 Results ................................................................................................................................................... 108 Discussion ............................................................................................................................................. 113 References ............................................................................................................................................. 120 Figures .................................................................................................................................................. 123 Tables .................................................................................................................................................... 126 Supplemental Material .........................................................................................................................