PROSTAGLANDINS IN PRIMATE OVULATORY FOLLICLES

灵长类动物排卵卵泡中的前列腺素

• 批准号: 6501792
• 负责人: Diane M Duffy
• 金额: $ 27.32万
• 依托单位: EASTERN VIRGINIA MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-02-15 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6501792
• 关键词: Macaca mulatta; angiogenesis factor; corpus luteum; enzyme inhibitors; enzyme linked immunosorbent assay; fatty acid biosynthesis; gene expression; genetic markers; gonadotropins; graafian follicles; granulosa cell; immunocytochemistry; in situ hybridization; messenger RNA; ovulation; polymerase chain reaction; progesterone; prostaglandin E; prostaglandin endoperoxide synthase; prostaglandin receptor; prostaglandins; radioimmunoassay; tissue /cell culture

DESCRIPTION: Preliminary studies conducted by the PI indicate that the ovulatory gonadotropin surge initiates prostaglandin (PG) production by the primate periovulatory follicle and the injection of a PG synthesis inhibitor directly into the follicle prevents follicle rupture without altering normal luteal progression. The proposed studies will address the hypothesis that PGs produced by the preovulatory follicle act locally to regulate specific molecular events related to follicular rupture, but not luteinization, of the primate follicle. Experiments are proposed to examine the locations and gonadotropin regulation of PG synthetic enzymes within the primate periovulatory follicle (Specific Aim 1). In vitro protocols will be used to determine if gonadotropin stimulation of granulosa and/or theca cells of the follicle is sufficient to initiate PG synthesis (Specific Aim 2). PG receptors will be localized to cells of the periovulatory follicle to determine the site(s) of PG action (specific Aim 3). PG-stimulated expression of molecular markers of follicle rupture (matrix metalloproteinases and their inhibitors [TIMPs], plasminogen activator and its inhibitor and luteinization (3beta hydroxydehydrogenase, steroidogenic acute regulatory protein [STAR], vascular endothelial growth factor (VEGF], angiopoietin-1 and 2) will be assessed (specific Aim 4). Immunocytochemistry and in situ hybridization will be used to localize PG synthetic enzymes and PG receptors to the cells of the preovulatory follicle. Gonadotropin regulation of mRNAs for PG synthetic enzymes will be determined by semi-quantitative RT-PCR based assays. PG production of cultured ovarian cells and tissues will be measured using EIA kits. In vitro expression of markers of follicular rupture/luteinization will be assessed using RT-PCR based assays for mRNAs, ELISA kits for enzyme proteins and angiogenic factors, and RIA for media progesterone. Intrafollicular injection of PG synthesis inhibitor will be used to determine if PGs regulate specific markers of follicular rupture and luteinization in vivo. These studies will likely demonstrate that the mid cycle gonadotropin surge initiates PG synthesis by primate ovarian follicle through the induction of specific enzymes and support the hypothesis that PGs act locally to trigger specific processes related to ovulation, but not luteinization, of the periovulatory follicle. This may provide insight into the mechanisms leading to infertility (e.g., leuteinized unruptured follice syndrome) and support the developoment of PG synthesis inhibitors of PG synthesis inhibitors for use as contraceptives.

描述：PI进行的初步研究表明， 排卵促性腺激素峰启动前列腺素（PG）的生产， 灵长类排卵期卵泡和注射PG合成抑制剂 直接进入卵泡防止卵泡破裂，而不改变正常的 黄体进展拟议的研究将解决的假设，PG 排卵前卵泡产生的激素局部调节特定的 与卵泡破裂有关的分子事件，但不是黄体化， 灵长类卵泡实验提出了检查的位置和 促性腺激素对灵长类PG合成酶的调节 排卵期卵泡（特定目标1）。体外方案将用于 确定是否促性腺激素刺激颗粒和/或卵泡膜细胞的 卵泡足以启动PG合成（特定目标2）。PG受体 将定位于排卵期卵泡的细胞，以确定 PG作用部位（具体目标3）。PG刺激的分子表达 卵泡破裂标志物（基质金属蛋白酶及其抑制剂 [TIMPs]、纤溶酶原激活剂及其抑制剂与黄体化（3 β 羟脱氢酶，类固醇生成急性调节蛋白[星星]，血管 将评估内皮生长因子（VEGF）、血管生成素-1和2 （具体目标4）。免疫细胞化学和原位杂交将用于 将PG合成酶和PG受体定位于排卵前细胞 毛囊PG合成酶的mRNA的促性腺激素调节将是 通过基于半定量RT-PCR的测定来确定。培养的PG产量 使用EIA试剂盒测量卵巢细胞和组织。体外表达 将使用RT-PCR评估卵泡破裂/黄体化的标志物 基于mRNA的测定，酶蛋白和血管生成因子的ELISA试剂盒， 放免法测定中孕酮。卵泡内注射PG合成 抑制剂将用于确定PG是否调节 卵泡破裂和体内黄体化。这些研究可能会 证明中期周期促性腺激素激增通过以下方式启动PG合成： 灵长类卵泡通过特异性酶的诱导和支持 假设PG在局部起作用以触发与以下相关的特定过程： 排卵期卵泡排卵，但不黄体化。这可以提供对导致不育的机制的深入了解（例如，亮氨酸化未破裂卵泡综合征），并支持PG合成抑制剂的开发，以用作避孕药。