Recent evidence in primates demonstrates that proliferation of granulosa cells is arrested within the first quarter of the periovulatory interval. However, specific markers of cell cycle progression, e.g., cyclin D2, are increased while levels of cell cycle suppressors such as p53 are reduced in concurrence with cell cycle arrest. This application will test the hypothesis that an ovulatory stimulus induces a transient burst of granulosa cell proliferation that is an essential feature of terminal differentiation and luteal formation in the primate. Experiments are planned to examine cell cycle characteristics of granulosa cells at early time points following a luteinizing dose of gonadotropin to granulosa cells in vitro and after an ovulatory stimulus to adult, female rhesus monkeys in vivo (aim 1). The functional consequences of the proliferative burst on terminal differentiation and luteinization of granulosa cells will be determined using in vitro protocols (aim 2). This application will further test the hypothesis that estrogen mediates the proliferative burst via regulation of key cell cycle components, and that early periovulatory estrogen action is essential to the formation of a functional corpus luteum (aim 3). The proposed studies will utilize an in vivo controlled ovarian stimulation model in which the differential effects of gonadotropins and steroids can be determined. Granulosa cells will be obtained from rhesus monkeys undergoing controlled ovarian stimulation prior to an ovulatory stimulus for in vitro experimentation. Levels of mRNA will be determined with real time RT-PCR, and protein levels and activity by western blot, gel shift, and kinase assays. Concentrations of steroid hormones will be measured by RIA. The proposed studies are expected to demonstrate that primate granulosa cells undergo a proliferative burst in response to the ovulatory stimulus that is an essential component of luteinization. It is expected that estrogen plays a heretofore unexpected role in driving the proliferative burst through the regulation of key cell cycle components. These studies will provide insight into the etiologies of ovarian cancer and certain kinds of infertility, as well as provide potential novel avenues for contraceptive agents.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD043358-02
Application #
6757247
Study Section
Reproductive Endocrinology Study Section (REN)
Program Officer
Taymans, Susan
Project Start
2003-07-01
Project End
2005-04-01
Budget Start
2004-05-01
Budget End
2005-04-01
Support Year
2
Fiscal Year
2004
Total Cost
$282,476
Indirect Cost
Name
Medical College of Georgia (MCG)
Department
Physiology
Type
Schools of Medicine
DUNS #
966668691
City
Augusta
State
GA
Country
United States
Zip Code
30912
Puttabyatappa, Muraly; Brogan, Rebecca S; Vandevoort, Catherine A et al. (2013) EGF-like ligands mediate progesterone's anti-apoptotic action on macaque granulosa cells. Biol Reprod 88:18
Brogan, Rebecca S; MacGibeny, Margaret; Mix, Scott et al. (2011) Dynamics of intra-follicular glucose during luteinization of macaque ovarian follicles. Mol Cell Endocrinol 332:189-95
Kolmakova, Antonina; Wang, Jiangxia; Brogan, Rebecca et al. (2010) Deficiency of scavenger receptor class B type I negatively affects progesterone secretion in human granulosa cells. Endocrinology 151:5519-27
Brogan, Rebecca S; Mix, Scott; Puttabyatappa, Muraly et al. (2010) Expression of the insulin-like growth factor and insulin systems in the luteinizing macaque ovarian follicle. Fertil Steril 93:1421-9
Puttabyatappa, Muraly; Vandevoort, Catharine A; Chaffin, Charles L (2010) hCG-induced down-regulation of PPAR? and liver X receptors promotes periovulatory progesterone synthesis by macaque granulosa cells. Endocrinology 151:5865-72
Cherian-Shaw, Mary; Puttabyatappa, Muraly; Greason, Erin et al. (2009) Expression of scavenger receptor-BI and low-density lipoprotein receptor and differential use of lipoproteins to support early steroidogenesis in luteinizing macaque granulosa cells. Endocrinology 150:957-65
Cannon, Jennifer D; Seekallu, Srinivas V; Vandevoort, Catherine A et al. (2009) Association of luteinizing hormone receptor gene expression with cell cycle progression in granulosa cells. Am J Physiol Endocrinol Metab 296:E1392-9
Nyholt de Prada, Jenna K; Lee, Young S; Latham, Keith E et al. (2009) Role for cumulus cell-produced EGF-like ligands during primate oocyte maturation in vitro. Am J Physiol Endocrinol Metab 296:E1049-58
de Prada, Jenna K Nyholt; Hill, Dana L; Chaffin, Charles L et al. (2009) Nuclear maturation and structural components of nonhuman primate cumulus-oocyte complexes during in vivo and in vitro maturation. Fertil Steril 91:2043-50
Cannon, Jennifer D; Cherian-Shaw, Mary; Lovekamp-Swan, Tara et al. (2007) Granulosa cell expression of G1/S phase cyclins and cyclin-dependent kinases in PMSG-induced follicle growth. Mol Cell Endocrinol 264:6-15

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