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Steroid-Dependent Regulation of the Oviduct: a Cross-Species Transcriptomal Analysis

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Steroid-Dependent Regulation of the Oviduct: a Cross-Species Transcriptomal Analysis University of Kentucky UKnowledge Theses and Dissertations--Animal and Food Sciences Animal and Food Sciences 2015 Steroid-dependent regulation of the oviduct: A cross-species transcriptomal analysis Katheryn L. Cerny University of Kentucky, [email protected] Right click to open a feedback form in a new tab to let us know how this document benefits ou.y Recommended Citation Cerny, Katheryn L., "Steroid-dependent regulation of the oviduct: A cross-species transcriptomal analysis" (2015). Theses and Dissertations--Animal and Food Sciences. 49. https://uknowledge.uky.edu/animalsci_etds/49 This Doctoral Dissertation is brought to you for free and open access by the Animal and Food Sciences at UKnowledge. It has been accepted for inclusion in Theses and Dissertations--Animal and Food Sciences by an authorized administrator of UKnowledge. For more information, please contact [email protected]. STUDENT AGREEMENT: I represent that my thesis or dissertation and abstract are my original work. Proper attribution has been given to all outside sources. I understand that I am solely responsible for obtaining any needed copyright permissions. I have obtained needed written permission statement(s) from the owner(s) of each third-party copyrighted matter to be included in my work, allowing electronic distribution (if such use is not permitted by the fair use doctrine) which will be submitted to UKnowledge as Additional File. I hereby grant to The University of Kentucky and its agents the irrevocable, non-exclusive, and royalty-free license to archive and make accessible my work in whole or in part in all forms of media, now or hereafter known. I agree that the document mentioned above may be made available immediately for worldwide access unless an embargo applies. I retain all other ownership rights to the copyright of my work. I also retain the right to use in future works (such as articles or books) all or part of my work. I understand that I am free to register the copyright to my work. REVIEW, APPROVAL AND ACCEPTANCE The document mentioned above has been reviewed and accepted by the student’s advisor, on behalf of the advisory committee, and by the Director of Graduate Studies (DGS), on behalf of the program; we verify that this is the final, approved version of the student’s thesis including all changes required by the advisory committee. The undersigned agree to abide by the statements above. Katheryn L. Cerny, Student Dr. P.J. Bridges, Major Professor Dr. D.L. Harmon, Director of Graduate Studies STEROID-DEPENDENT REGULATION OF THE OVIDUCT: A CROSS-SPECIES TRANSCRIPTOMAL ANALYSIS ________________________________________ DISSERTATION _________________________________________ A dissertation submitted in partial fulfillment of the Requirements for the degree of Doctor of Philosophy in the College of Agriculture at the University of Kentucky By Katheryn Leigh Cerny Lexington, Kentucky Director: Dr. Phillip J. Bridges, Ph.D. Assistant Professor Lexington, Kentucky Copyright © Katheryn Leigh Cerny, 2015 ABSTRACT OF DISSERTATION STEROID-DEPENDENT REGULATION OF THE OVIDUCT: A CROSS-SPECIES TRANSCRIPTOMAL ANALYSIS Reproductive success depends on a functional oviduct for gamete storage, maturation, fertilization, and early-conceptus development. The ovarian-derived sex steroids estradiol and progesterone are known to affect functionality of the oviduct. Advances in microarray and NanoString technology allow for gene expression analysis to increase understanding of processes critical for fertility. Studies were conducted to investigate mechanisms regulating oviductal function in cattle and mice by using the Bovine Gene 1.0 ST array and the Mouse Gene 430-2.0 arrays (Affymetrix Inc., CA), respectively. For the first study, oviducts were collected from heifers assigned to luteal or follicular phase groups. In the second study oviducts were collected from immature mice with a global deletion of estrogen receptor-1 (ESR1) and their wild-type littermates at 23 days of age or 48 hr after treatment with 5 IU of PMSG. Following microarray hybridization, the resulting datasets were analyzed using Partek Genomics Suite 6.6 (Partek Inc., MO). The results of the first two studies illustrated a dynamic hormonal regulation of the oviductal epithelium and revealed the identity of novel genes affecting fertility in cattle and gave us insights into the genes regulated by estrogen and ESR1 in mice. Many genes identified as differentially regulated are believed to play an integral role in the regulation of oviductal inflammation. Therefore, the objective of the third study was to test the hypothesis that intraperitoneal administration of E. Coli-derived lipopolysaccharide induces the expression of inflammatory mRNAs in the mouse oviduct. Mice were treated with 0, 2 μg or 10 μg of LPS from E. Coli. and killed 24 h later. Oviducts were collected for determination of inflammatory gene expression by a targeted NanoString approach using the nCounter GX Mouse Inflammation Kit (NanoString Technologies, Wa). Results indicate that systemic treatment with LPS induces inflammation in the oviducts of mice and provides evidence of a repeatable animal model of oviductal inflammation. Overall, data from these studies extends our knowledge of the mechanisms regulating oviductal functions and immune response, as well as identified target molecules and processes to improve production animal and human fertility. Keywords: Oviduct, Steroid hormones, LPS-induced inflammation, Tubal-factor infertility, immune response Katheryn L. Cerny Student’s Signature 7/21/2015 Date STEROID-DEPENDENT REGULATION OF THE OVIDUCT: A CROSS-SPECIES TRANSCRIPTOMAL ANALYSIS By Katheryn Leigh Cerny Dr. P.J. Bridges Director of Dissertation Dr. J.C. Matthews Co-Chair of Dissertation Dr. D.L. Harmon Director of Graduate Studies 7/21/2015 Date ACKNOWLEDGEMENTS Earning a Ph.D. has been a lifelong goal and I am fortunate to have had an inspiring group of advisors along the way. First and foremost I would like to thank Dr. P.J. Bridges, my Ph.D. advisor and chairman of my doctoral committee for guiding me through my graduate career. I first met Dr. Bridges as an equine professional looking to expand my experiences in research to other species. It was my lucky day when Dr. Bridges offered me a position as a graduate research assistant in his lab where I could learn valuable lab techniques applicable to reproductive physiology and work with cattle and mice for my Ph.D. Through all the twists and turns of graduate school I could always count on Dr. Bridges to help guide my direction. I would also like to express my appreciation and thanks to my doctoral committee members: Co-chair Dr. J.C. Matthews, Dr. L. Anderson, Dr. K. McDowell, and Dr. B.A. Ball for their advice and support during my Ph.D. projects and allowing me to grow as a research scientist. I would especially like to acknowledge my parents, Bridget and Max Cerny for their continuous encouragement and support in achieving my goals. I am extremely grateful for the sacrifices they have made throughout my life and I consider myself an exceptionally fortunate person. iii TABLE OF CONTENTS ACKNOWLEDGEMENTS ...................................................................................................................... iii LIST OF TABLES ..................................................................................................................................... vi LIST OF FIGURES .................................................................................................................................. vii FREQUENTLY USED ABBREVIATIONS ....................................................................................... viii CHAPTER 1. INTRODUCTION............................................................................................................. 1 CHAPTER 2. A REVIEW OF THE LITERATURE ........................................................................... 3 Structure of the Oviduct .............................................................................................................. 3 Steroidal Regulation of the Oviduct ........................................................................................ 4 Gamete Storage and Maturation .............................................................................................. 7 Fertilization and Conceptus Development .........................................................................10 Oviduct Epithelial Cell Secretions .........................................................................................11 Inflammatory Response in the Oviduct ...............................................................................13 CHAPTER 3. STATEMENT OF THE PROBLEM ..........................................................................16 CHAPTER 4. A TRANSCRIPTOMAL ANALYSIS OF OVIDUCTAL EPITHELIAL CELLS COLLECTED DURING THE FOLLICULAR VERSUS LUTEAL PHASE IN CATTLE ...........19 Abstract ............................................................................................................................................19 Introduction ...................................................................................................................................21 Methods ...........................................................................................................................................23 Results and Discussion ..............................................................................................................29
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