Gametogenesis in females is controlled by follicle-stimulating hormone (FSH) from the pituitary, which is, in turn, regulated by gonadotropin-releasing hormone (GnRH) from the hypothalamus. GnRH regulates FSH synthesis through induction of the immediate early gene, c-Fos, which, upon heterodimerization with c-Jun, creates an AP-1 transcription factor that binds the promoter of the FSH 2-subunit gene. The overall goal of this application is to ascertain the mechanism of GnRH induction of c-Fos and to elucidate the role of AP-1 in the gonadotrope in vivo. AP-1 is the only factor known to be involved in the induction of the FSH 2-subunit gene by GnRH. c-Fos, the most highly GnRH-induced gene in the gonadotrope, is the most regulated AP-1 isoform, while c-Jun expression varies less with hormone treatment. The molecular mechanism of c-Fos induction by GnRH in the gonadotrope cell is not known. Neither has a role for AP-1 in reproduction in vivo been determined. Herein, I will utilize novel approaches to delineate the mechanisms underlying the regulation of c- Fos expression by GnRH. In the first aim, I will determine the molecular mechanism of c-Fos induction by GnRH. In the second aim, I will determine the regulation of protein stability and degradation, since c-Fos is an immediate-early gene with a very unstable message and labile protein, and its cellular concentration is controlled on the level of turnover, as much as on the level of expression. In the third aim of this proposal, I will determine the function of AP-1 in reproduction in vivo with the use of genetically modified animals. First, I will assess reproductive function and gonadotropin expression in c-Fos deficient animals and then, as complementary approach, use mice that lack c-Jun specifically in the pituitary gonadotrope. Jun isoforms are obligatory heterodimeric partners for Fos isoforms, whose interaction creates an AP-1 transcription factor that binds DNA. c-Jun, in particular, is essential for development, since knock-out animals die in utero, and with the use of CRE-lox system, I will analyze the function of c-Jun in gonadotrope in vivo. Results obtained from these studies will make substantial contributions to our understanding of the molecular mechanisms that affect gene expression in the gonadotrope and provide insight into the physiology and pathophysiology of the mammalian reproductive system. I propose to expand our understanding of molecular mechanisms of GnRH induction of c- Fos gene in the gonadotrope in a cell culture model, which will lead to a better understanding of GnRH regulation of gene expression in the pituitary, and the function of AP-1 in vivo, utilizing genetically modified mice.

Public Health Relevance

Fertility in mammals is regulated primarily by gonadotropin-releasing hormone effect on the gonadotrope cell population in the pituitary gland, through modulation of gonadotropin gene expression and secretion. Gonadotropins, follicle-stimulating hormone and luteinizing hormone, work on the gonads to stimulate gametogenesis and steroidogenesis. This proposal will address the molecular mechanism of gene regulation by GnRH in the gonadotrope cell.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
7R01HD057549-05
Application #
8452618
Study Section
Integrative and Clinical Endocrinology and Reproduction Study Section (ICER)
Program Officer
Lamar, Charisee A
Project Start
2009-04-01
Project End
2015-03-31
Budget Start
2013-04-01
Budget End
2015-03-31
Support Year
5
Fiscal Year
2013
Total Cost
$276,813
Indirect Cost
$92,911
Name
University of California Riverside
Department
None
Type
Schools of Medicine
DUNS #
627797426
City
Riverside
State
CA
Country
United States
Zip Code
92521
Glidewell-Kenney, Christine A; Trang, Crystal; Shao, Paul P et al. (2014) Neurokinin B induces c-fos transcription via protein kinase C and activation of serum response factor and Elk-1 in immortalized GnRH neurons. Endocrinology 155:3909-19
Roybal, Lacey L; Hambarchyan, Arpi; Meadows, Jason D et al. (2014) Roles of binding elements, FOXL2 domains, and interactions with cJUN and SMADs in regulation of FSH?. Mol Endocrinol 28:1640-55
Witham, Emily A; Meadows, Jason D; Hoffmann, Hanne M et al. (2013) Kisspeptin regulates gonadotropin genes via immediate early gene induction in pituitary gonadotropes. Mol Endocrinol 27:1283-94
Lindaman, Lacey L; Yeh, Debra M; Xie, Changchuan et al. (2013) Phosphorylation of ATF2 and interaction with NFY induces c-Jun in the gonadotrope. Mol Cell Endocrinol 365:316-26
Reddy, Gaddameedi R; Xie, Changchuan; Lindaman, Lacey L et al. (2013) GnRH increases c-Fos half-life contributing to higher FSH? induction. Mol Endocrinol 27:253-65
Ely, Heather A; Mellon, Pamela L; Coss, Djurdjica (2011) GnRH induces the c-Fos gene via phosphorylation of SRF by the calcium/calmodulin kinase II pathway. Mol Endocrinol 25:669-80
Breen, Kellie M; Thackray, Varykina G; Coss, Djurdjica et al. (2010) Runt-related transcription factors impair activin induction of the follicle-stimulating hormone {beta}-subunit gene. Endocrinology 151:2669-80
Coss, Djurdjica; Mellon, Pamela L; Thackray, Varykina G (2010) A FoxL in the Smad house: activin regulation of FSH. Trends Endocrinol Metab 21:562-8
Thackray, Varykina G; Mellon, Pamela L; Coss, Djurdjica (2010) Hormones in synergy: regulation of the pituitary gonadotropin genes. Mol Cell Endocrinol 314:192-203
Corpuz, Patrick S; Lindaman, Lacey L; Mellon, Pamela L et al. (2010) FoxL2 Is required for activin induction of the mouse and human follicle-stimulating hormone beta-subunit genes. Mol Endocrinol 24:1037-51

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