PROSTAGLANDIN RECEPTOR REGULATION IN THE CORPUS LUTEUM

前列腺素(PG)在调节卵巢功能中起着重要作用 包括排卵、黄体生成和黄体退化的关键方面。 尽管在生殖生理学中扮演着核心角色，但时间和 PG受体的细胞表达模式，以及 荷尔蒙对这种表达的调节，还没有定义。与 前列腺素E和前列腺素F/2α各亚型基因的最新克隆 受体，对这些受体的蛋白质和mRNA的分析是 有可能。我们已经验证了高度敏感和定量的分析方法 前列腺素F/2α受体的mRNA和蛋白及其检测 PGF/2α反应性。这项申请中提出的研究是 旨在评估参与表达该基因的机制 前列腺素F/2α受体与黄体生成素的反应性 黄体溶解。我们对这些研究的工作假设是，黄体生成素 激增或黄体化过程的其他方面，会导致 前列腺素F/2α受体的表达与细胞内PG反应 因此，建立排卵的细胞机制的途径， 黄体化和黄体退化。具体目标 包括： 1)表征在体内表达的时间模式 PGF/2α受体在卵泡晚期和黄体早期发育中的作用 2)确定cAMP依赖的蛋白激酶、孕酮、 PGF/2α分化过程中的从头蛋白合成 培养的滤泡细胞的反应性。 3)确定Forskolin或Forsklin是否能诱导小黄体细胞 绒毛膜促性腺激素表达FP受体，这是一种指示大黄体的特性 细胞表型。确定参与的细胞内效应系统 前列腺素F/2α对前列腺素H合成酶-2基因的刺激和抑制作用 PGF/2α受体的mRNA的表达。 4)确定早、中期黄体库对 前列腺素F/2α对前列腺素F/2α受体分泌的下调作用 催产素的表达，诱导PGHS-2的mRNA表达，并抑制其表达 3β-羟基类固醇脱氢酶基因的表达。 这些研究将促进我们对生理、细胞内、 以及调控基因表达的分子机制。 前列腺素F/2α受体的细胞内反应途径。这将是 不仅提高了我们目前对生殖生理学的认识，而且 将允许合理开发避孕药具或不孕不育 针对卵巢中前列腺素作用的治疗。

Prostaglandins (PG) have an essential role in regulating ovarian function including key aspects of ovulation, luteinization, and luteal regression. Despite this central role in reproductive physiology the temporal and cellular patterns of expression for the PG receptors, as well as the hormonal regulation of this expression, have not been defined. With the recent cloning of the cDNAs for the various subtypes of PGE and PGF/2alpha receptors, an analysis of both the protein and mRNA for these receptors is possible. We have validated highly sensitive and quantitative assays for the PGF/2alpha receptor mRNA and protein as well as the measuring PGF/2alpha responsiveness. The research proposed in this application is designed to evaluate the mechanisms involved in expression of the PGF/2alpha receptors and responsiveness during luteinization and luteolysis. Our working hypotheses for these studies is that the LH surge, or other aspects of the luteinization process, induce the expression of PGF/2alpha receptors and PG-responsive intracellular pathways, thus, establishing the cellular mechanisms for ovulation, luteinization, and regression of the corpus luteum. The Specific Aims are: 1) Characterize the temporal pattern for the in vivo expression of PGF/2alpha receptors during late follicular and early luteal development. 2) Determine the role of cAMP-dependent protein kinase, progesterone, and de novo protein synthesis in the differentiation of PGF/2alpha responsiveness in cultured follicular cells. 3) Determine whether small luteal cells can be induced by forskolin or hCG to express FP receptors, a property indicative of the large luteal cell phenotype. Determine the intracellular effector systems involved in PGF/2alpha-induced stimulation of mRNA for PGH synthase-2 and inhibition of mRNA for the PGF/2alpha receptor. 4) Determine the response of early and mid-cycle corpora lutea to PGF/2alpha in terms of down regulation of PGF/2alpha receptors, secretion of oxytocin, induction of mRNA for PGHS-2, and inhibition of expression of mRNA for 3beta-Hydroxysteroid Dehydrogenase. These studies will advance our knowledge of physiologic, intracellular, and molecular mechanisms regulating the expression of mRNA, protein, and intracellular response pathways for the PGF/2alpha receptor. This will not only advance our current knowledge of reproductive physiology, but will allow for the rational development of contraceptives or infertility treatments that target prostaglandin action in the ovary.