PROSTAGLANDINS IN PRIMATE OVULATORY FOLLICLES

灵长类动物排卵卵泡中的前列腺素

• 批准号: 6499157
• 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/6499157
• 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：由该署进行的初步研究显示， 排卵性促性腺激素激增通过促性腺激素释放 灵长类围排卵泡与PG合成抑制剂的注射 直接进入卵泡可防止卵泡破裂而不改变正常状态 黄体进展。拟议的研究将解决以下假设：PGs 由排卵前卵泡产生的局部作用调节特定的 与卵泡破裂有关的分子事件，但与黄体化无关 灵长类卵泡。建议进行实验，以检查位置和 促性腺激素对灵长类体内PG合成酶的调节 排卵周卵泡(特定目标1)。体外方案将被用来 确定促性腺激素对卵巢颗粒细胞和/或卵泡膜细胞的刺激作用 卵泡足以启动PG合成(特定目标2)。PG受体 将定位于排卵周围卵泡的细胞以确定 PG动作现场(S)(具体目标3)。PG刺激的分子表达 卵泡破裂的标志物(基质金属蛋白酶及其抑制物 [TIMPs]、纤溶酶原激活物及其抑制物与黄体化(3beta 羟基脱氢酶，类固醇合成急性调节蛋白[STAR]，血管 将评估血管内皮生长因子(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 PG合成酶和PG受体在排卵前期细胞中的定位 毛囊。促性腺激素对PG合成酶mRNAs的调节作用 通过基于半定量RT-PCR的检测。发酵生产PG的研究进展 卵巢细胞和组织将使用EIA试剂盒进行测量。体外表达 卵泡破裂/黄体化的标记物将使用RT-PCR进行评估 基于对mRNAs的检测，酶蛋白和血管生成因子的酶联免疫吸附试验试剂盒， 媒体黄体酮的放射免疫分析。卵泡内注射PG合成 抑制剂将被用来确定PGs是否调节特定的标志物 体内卵泡破裂和黄体化。这些研究很可能 证明周期中期促性腺激素激增通过以下途径启动PG合成 灵长类卵泡通过特定酶的诱导和支持 假设PG在本地行动以触发与以下相关的特定过程 排卵期卵泡的排卵，但不是黄体化。这可能有助于深入了解导致不孕症的机制(例如，白化未破裂卵泡综合征)，并支持PG合成抑制剂的开发作为避孕用药。