University of New Hampshire University of New Hampshire Scholars' Repository Doctoral Dissertations Student Scholarship Winter 2005 On the neuroendocrine regulation of reproduction: Functional characterization and kinetic studies of the lamprey gonadotropin -releasing hormone receptor and cloning and analysis of the cDNA encoding lamprey gonadotropin-releasing hormone-III Matthew Ren Silver University of New Hampshire, Durham Follow this and additional works at: https://scholars.unh.edu/dissertation Recommended Citation Silver, Matthew Ren, "On the neuroendocrine regulation of reproduction: Functional characterization and kinetic studies of the lamprey gonadotropin -releasing hormone receptor and cloning and analysis of the cDNA encoding lamprey gonadotropin-releasing hormone-III" (2005). Doctoral Dissertations. 309. https://scholars.unh.edu/dissertation/309 This Dissertation is brought to you for free and open access by the Student Scholarship at University of New Hampshire Scholars' Repository. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of University of New Hampshire Scholars' Repository. For more information, please contact [email protected]. ON THE NEUROENDOCRINE REGULATION OF REPRODUCTION: FUNCTIONAL CHARACTERIZATION AND KINETIC STUDIES OF THE LAMRPEY GONADOTROPIN-RELEASING HORMONE RECEPTOR AND CLONING AND ANALYSIS OF THE cDNA ENCODING LAMPREY GONADOTROPIN-RELEASING HORMONE-III BY MATTHEW REN SILVER B.S., University of Connecticut, 2000 DISSERTATION Submitted to the University of New Hampshire in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy In Biochemistry December, 2005 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. UMI Number: 3198014 INFORMATION TO USERS The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleed-through, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. ® UMI UMI Microform 3198014 Copyright 2006 by ProQuest Information and Learning Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. ProQuest Information and Learning Company 300 North Zeeb Road P.O. Box 1346 Ann Arbor, Ml 48106-1346 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. This dissertation has been examined and approved. 2 k LLL<a Dissertation Director, Dr. Stacia A. Sower Interim Associate Dean of Research of the College of Life Sciences and Agriculture, Professor of Biochemistry and Molecular Biology Dr. William R. Dean, College of Life Sciences and Agriculture, Professor of Biochemistry and Molecular Biology Dr. William A. Condon Professor of Animal and Nutritional Sciences Dr. William K. Thomas Associate Professor of Biochemistry and Molecular Biology Dr. Karen L. Carleton Research Associate Professor of Zoology M(Xi 1*3,? fn C Date Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. DEDICATION To Molly iii Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ACKNOWLEDGEMENTS This research and my academic development would never have been possible if it were not for many people. Without question, all of my achievements were made possible directly as a result of Professor Stacia Sower’s guidance and support. I cannot fully express my gratitude for Stacia’s patience and confidence to let me develop my projects independently and her willingness and flexibility to allow me to branch out and pursue my interests. Most important of all I want to express my appreciation for Stacia’s consistent positive feedback, endorsement and promotion. Finally, I thank Stacia for occasionally letting me win at golf. I would also like to thank all of the members of my committee, Drs. William Trumble, William Condon, Kelley Thomas and Karen Carleton; all of whom provided invaluable insight and advice. I would lastly like to thank Professor Andy Laudano and the late Professor Charles Warren for their advice and assistance in the early stages of cell culture and transfection development. Non-academic support is just as important as the aforementioned sort, of which I received the best kind, love and understanding, from my wife, Molly. Molly’s strengths are many; however she particularly shines when it comes to organization, neatness and the prompt and thorough execution and completion of duties. If it were not for her I would still be working on putting together my graduate school application packets to this date. I thank Molly for many things, but most of all for always being there. iv Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. As luck would have it, or fate or maybe foul chance, on my first day at UNH I nervously strolled into Joan Holt’s brutal radiation safety class and crossed paths with Gimli in a black Harley shirt. At the time I never would have guessed that I just met my work wife. He is Scott Kavanaugh, master of the morning, wise in Sower Lab lore (and other), hagfish slayer and a true friend. Scott made this experience fun, interesting and survivable. Thank you. I next met Adam Root, first by style in biochemistry lecture and then in person in the Sower lab. Adam’s upbeat and positive energy makes him a fun person to be around. He introduced me to the world of Flynn and Clancy, and can out eat just about anybody. Most of all, I thank Adam for his assistance and reference during my application. And then there was Nathaniel. Nathaniel Nucci that is, extremely smart and handy with the golden shovel, but not such a great gym partner. Nathaniel cloned the full length lamprey GnRH receptor, which paved the way for the research constituting the bulk of this dissertation. I appreciate this. Furthermore, his generosity during the publication process and in friendship deserves my thanks and praise. I would also like to thank all of my other cohorts, Sower lab members both past and present, including Mihael Freamat, Dr. Kara Lee, Dr. Mickie Powerll, Cari Boum, Emily Violette, Jocelyn Sanford, Jenny Glieco, Greg Blaisdell, Gregory Jerome, Ben Lyons, Jason Dahlstrom, Mark Santos, Bumie Schultz, Mike Baron, Geoff Bushold, Kazu Okamoto and Emily Lafiandra. Finally, I thank my family: my mother, April, father and step-mother, Joe and Emily, and brothers and sister, Ben, Emily and Dan. I greatly appreciate all of your love and support, from April 13th, 1977 to present. v Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. TABLE OF CONTENTS DEDICATION iii ACKNOWLEDGEMENTS................................................................................................. iv LIST OF FIGURES............................................................................................................... ix LIST OF TABLES................................................................................................................. xi ABSTRACT.......................................................................................................................... xii CHAPTER PAGE I. BACKGROUND AND SIGNIFICANCE.................................................................... 1 Gonadotropin-Releasing Hormone ............................................................................2 GnRH Development: Embryonic Origin and Cellular Migration ............................8 Phylogeny of the GnRH Family .................................................................................. 10 GnRH: From Mature Peptide to Gene.........................................................................12 GnRH Neuron Regulation ........................................................................................... 15 Clinical Applications of GnRH and its Analogs ........................................................ 21 Gonadotropin-Releasing Hormone Receptor ..............................................................23 Vertebrates Express Multiple GnRH Receptors ........................................................ 26 Structure and Regulation of the GnRH Receptor Gene ............................................ 29 GnRH Receptor Structure ............................................................................................ 32 vi Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. GPCR Activation ..........................................................................................................34 Signaling Systems of the GnRH Receptor ..................................................................37 Regulation of the GnRH Receptor: Functional Significance of the C-Terminal Tail.........................................................................................................40 Agnathans ..................................................................................................................... 45 Petromyzoniformes .......................................................................................................46 Lamprey Ecology ..........................................................................................................49