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Neonatal Growth Measurements, Umbilical Concentrations, And Clemson University TigerPrints All Theses Theses 12-2017 Neonatal Growth Measurements, Umbilical Concentrations, and Differential Gene Expression in the Placenta of Swine in Relation to Placental Efficiency Shanice Katelin Krombeen Clemson University Follow this and additional works at: https://tigerprints.clemson.edu/all_theses Recommended Citation Krombeen, Shanice Katelin, "Neonatal Growth Measurements, Umbilical Concentrations, and Differential Gene Expression in the Placenta of Swine in Relation to Placental Efficiency" (2017). All Theses. 2789. https://tigerprints.clemson.edu/all_theses/2789 This Thesis is brought to you for free and open access by the Theses at TigerPrints. It has been accepted for inclusion in All Theses by an authorized administrator of TigerPrints. For more information, please contact [email protected]. NEONATAL GROWTH MEASUREMENTS, UMBILICAL CONCENTRATIONS, AND DIFFERENTIAL GENE EXPRESSION IN THE PLACENTA OF SWINE IN RELATION TO PLACENTAL EFFICIENCY A Thesis Presented to the Graduate School of Clemson University In Partial Fulfillment of the Requirements for the Degree Master of Science Animal and Veterinary Science by Shanice Katelin Krombeen December 2017 Accepted by: Dr. Tiffany A. Wilmoth, Committee Chair Dr. Julia L. Sharp Dr. Christopher A. Saski i ABSTRACT Placental efficiency (PE) describes the relationship between placental and fetal weights and is defined as fetal weight divided by placental weight. Within pig litters, PE can vary drastically, resulting in similarly sized pigs associated with very different placenta, up to a 50% weight difference. However, the means enabling the smaller placenta to grow a similarly sized littermate is unknown. The main objectives of this study were to (1) determine the concentration of glucose and cortisol in venous umbilical blood of pigs at birth and evaluate if a relationship existed between concentration and PE, and (2) determine the expression level of genes in placental and associated endometrial tissues of high PE and low PE feto-placental units. For the first objective, terminal Yorkshire crossed pigs (n = 15) were monitored during farrowing to tag umbilical cords and ear notch pigs to ensure feto-placental units were properly matched and PE could be calculated. Neonatal growth measurements were taken to compare high PE and low PE feto-placental units. Umbilical blood samples were taken for quantification of glucose and cortisol. The basis of high PE, a smaller but more efficient placenta, was confirmed as placental weight was reduced, but fetal weight was not different in the high PE group. Given that birth weight, crown-rump length, girth, and body weights up to day 21 did not differ by PE, the survival and postnatal growth performance of pigs grown of high PE placentas should not be reduced. Furthermore, glucose and cortisol concentrations did not differ by PE suggesting high PE placentas have some other means enabling the transport of similar nutrient and hormone quantities despite a reduction in size. For the second objective, maternal line gilts (n = 8) were ovario-hysterectomized on day 95 of gestation ii to obtain corresponding placental and endometrial samples from each feto-placental unit. PE was calculated to identify the most efficient and least efficient unit in each litter; placental and endometrial samples from these units formed the high PE and low PE comparison groups. RNA sequencing was performed to identify differentially expressed genes (DEG) in high PE compared to low PE placental and endometrial samples. In the placenta, 214 DEG were identified, while zero DEG were identified in the endometrium. Of those DEG in the placenta, 103 were upregulated and 111 were downregulated. Although a portion of the DEG identified in the pig placenta encoded nutrient transporters and gene products with angiogenic or growth factor activity, DEG with alternative functions were also identified, indicating the complexity of the relationship between placental and fetal weights. Overall the results of this study provide new insights into the regulation of PE; however, further research is required to make PE production applicable. iii DEDICATION I’d like to give a special thanks to Dr. Tiffany Wilmoth, my major advisor. Your expertise and advice have been invaluable to me. Thank you for guiding me through this journey! I’d also like to thank my other committee members, Dr. Christopher Saski and Dr. Julia Sharp. Thank you for your time, advice, and assistance. Furthermore, I’d like to thank all of the undergraduate students who assisted throughout the various stages of my research project, especially Leo Krick and Savannah Richmond. I’d also like to thank my fellow graduate student Jenny Presgraves for proofreading sections of my writing. Likewise, my academic endeavors would not have been possible without my support system. Thank you Mom and Dad for supporting me and giving me the opportunity to pursue a higher degree. Mom, thank you for always reminding me that I would make it through the stressful days. Dad, thank you for giving me the time I needed to find my passion. Vanessa, thank you for being an awesome sister and providing a fun escape from school. Lastly, thank you Chris McGuffin for being my rock through it all. iv ACKNOWLEDGMENTS I’d like to acknowledge the Clemson University Genomics & Computational Laboratory for their collaboration on the RNA sequencing portion of this project. Specifically, I’d like to thank Jeanice Troutman and Barbra Blackmon for their help in the genetics lab, and Dr. Christopher Saski, Vijay Shankar, and Rooksie Noorai for their analyses and explanations of the RNA sequencing results. Likewise, I’d like to acknowledge Dr. Julia Sharp for her assistance with the statistical analyses. I’d also like to thank Dr. Matthew Wilson at West Virginia University for providing the porcine placental and endometrial samples used for RNA sequencing. v TABLE OF CONTENTS Page TITLE PAGE .................................................................................................................... i ABSTRACT ..................................................................................................................... ii DEDICATION ................................................................................................................ iv ACKNOWLEDGMENTS ............................................................................................... v LIST OF TABLES ........................................................................................................ viii LIST OF FIGURES ......................................................................................................... x CHAPTER I. LITERATURE REVIEW .............................................................................. 1 Introduction .............................................................................................. 1 Development of the Pig Placenta ............................................................. 3 Classification of the Pig Placenta ............................................................ 8 Placental Vascularity ............................................................................... 9 Function of the Placenta ......................................................................... 10 Nutrient Transport and Utilization ......................................................... 12 Hormones ............................................................................................... 23 Variation in Litter Size ........................................................................... 25 Placental Efficiency in Pigs .................................................................. 28 Genetics .................................................................................................. 32 Conclusion ............................................................................................. 36 II. PLACENTAL EFFICIENCY IN RELATION TO NEONATAL GROWTH MEASUREMENTS, AND THE CONCENTRATION OF GLUCOSE AND CORTISOL IN THE UMBILICAL BLOOD OF PIGS .................................................................................. 38 Abstract .................................................................................................. 38 Introduction ............................................................................................ 39 Material and Methods ............................................................................ 41 Results .................................................................................................... 45 Discussion .............................................................................................. 56 vi Table of Contents (Continued) Page III. THE IDENTIFICATION OF DIFFERENTIALLY EXPRESSED GENES BETWEEN HIGH PLACENTAL EFFICIENCY AND LOW PLACENTAL EFFICIENCY PLACENTAS IN SWINE ...................................................................... 65 Abstract .................................................................................................. 65 Introduction ............................................................................................ 67 Materials and Methods ........................................................................... 69 Results .................................................................................................... 75 Discussion ............................................................................................ 114 IV. CONCLUSION .........................................................................................
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