Prostaglandin Receptors in Primate Ovulatory Follicles

灵长类动物排卵卵泡中的前列腺素受体

基本信息

批准号：
7842230
负责人：
Diane M Duffy
金额：
$ 7.04万
依托单位：
EASTERN VIRGINIA MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-08-09 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7842230
关键词：
Adverse effects Agonist Antral Attention Basement membrane Cell Culture Techniques Cell physiology Cells Contraceptive Agents Data Detection Development Dinoprostone Dose Event Failure Fertilization G-Protein-Coupled Receptors Gases Generations Gonadotropins Human In Vitro Infertility Injection of therapeutic agent Investigation Laboratories Lead Luteinizing Hormone Macaca Macaca fascicularis Mediating Mediator of activation protein Messenger RNA Monkeys Oocytes Ovarian Ovarian Hyperstimulation Syndrome Ovarian Stimulations Ovarian Tissue Ovary Ovulation Peptide Hydrolases Play Primates Process Production Progesterone Prostaglandin Receptor Prostaglandins Protein Kinase Proteins Reproduction Research Personnel Risk Role Rupture Signal Pathway Signal Transduction Steroid biosynthesis Techniques Testing Tissues granulosa cell in vivo laser capture microdissection novel oocyte maturation prevent programs receptor research study response steroid hormone success transcription factor

项目摘要

Ovulation is essential for successful reproduction. The ovulatory luteinizing hormone surge stimulates periovulatory follicles to produce prostaglandin E2 (PGE2), which is required for expansion of the cumulus granulosa cells, follicle rupture, and oocyte release. Blockade of PGE2 production causes ovulation failure, supporting investigation of PGE2 as a treatment for infertility as well as inhibition of PGE2 synthesis/action as a potential contraceptive. To unravel the mechanism by which PGE2 stimulates periovulatory events, this proposal puts forth the overall hypothesis that each of the four PGE2 (EP) receptors has a unique function in ovulation. First, we will determine which EP receptors are expressed by functionally-distinct subpopulations of granulosa cells within periovulatory follicles. Using laser capture microdissection as well as immunofluorescent EP detection in monkey ovarian tissue sections, we will identify the EP receptor(s) present in cumulus, basal mural, and antral mural granulosa cells as well as granulosa cells located at the follicle apex. These subpopulations each play a unique role in ovulation, and we anticipate that each subpopulation expressesa unique subset of all EP receptors. Second, we will demonstrate that each EP receptor mediates a subset of all PGE2-stimulated periovulatory processes. PGE2 is known to stimulate granulosa cell progesterone production, cumulus expansion, and follicle rupture. Granulosa cell cultures as well as intrafollicular injection in macaques will be utilized to demonstrate that agonists specific for each EP receptor stimulate some, but not all, of these essential PGE2-mediated granulosa cell functions. Finally, we will determine which EP receptors can transduce the PGE2 signal in granulosa cells of primate periovulatory follicles. Granulosa cells will be treated with agonists specific for each of the four EP receptors, and intracellular signals generated via these G protein coupled receptors will be assessed. Each EP receptor will likely respond to agonist stimulation with a unique intracellular response. The proposed studies will likely demonstrate that each EP receptor mediates a specific subset of the total periovulatory responseto PGE2 within the primate follicle. These data will facilitate a targeted pharmacological approach to selectively stimulate or inhibit essential features of the ovulatory process and may ultimately lead to the development of new treatments for infertility and novel contraceptive options.

排卵对于成功繁殖至关重要。排卵黄体生成激素激增刺激排卵期卵泡产生前列腺素 E2 (PGE2)，这是卵丘扩张所必需的颗粒细胞、卵泡破裂和卵母细胞释放。 PGE2 产生受阻会导致排卵失败，支持 PGE2 作为不孕症治疗方法以及抑制 PGE2 合成/作用的研究作为一种潜在的避孕药。为了揭示 PGE2 刺激排卵期事件的机制，本研究该提案提出了总体假设，即四种 PGE2 (EP) 受体中的每一种在排卵。首先，我们将确定功能不同的亚群表达哪些 EP 受体排卵期卵泡内的颗粒细胞。使用激光捕获显微切割以及对猴卵巢组织切片进行免疫荧光 EP 检测，我们将鉴定 EP 受体存在于积云、基底壁和窦壁颗粒细胞以及位于卵泡顶端。这些亚群在排卵中各自发挥着独特的作用，我们预计每个亚群亚群表达所有 EP 受体的独特子集。其次，我们将证明每个 EP 受体介导所有 PGE2 刺激的排卵过程的一部分。 PGE2 已知可刺激颗粒细胞黄体酮产生、卵丘扩张和卵泡破裂。颗粒细胞培养物为将利用猕猴卵泡内注射来证明每个 EP 具有特异性的激动剂受体刺激部分但不是全部 PGE2 介导的颗粒细胞功能。最后，我们将确定哪些 EP 受体可以转导灵长类排卵期颗粒细胞中的 PGE2 信号毛囊。颗粒细胞将用四种 EP 受体特异性激动剂处理，并且将评估通过这些 G 蛋白偶联受体产生的细胞内信号。每个 EP 受体都会可能对激动剂刺激做出独特的细胞内反应。拟议的研究可能会证明每个 EP 受体介导 PGE2 总排卵反应的特定子集在灵长类动物的卵泡内。这些数据将有助于有针对性的药理学方法选择性地刺激或抑制排卵过程的基本特征，并可能最终导致发展不孕不育的新疗法和新的避孕选择。