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：
7481086
负责人：
Diane M Duffy
金额：
$ 34.24万
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/7481086
关键词：
Adverse effects Agonist Andro-Diane Antral Attention Basement membrane Cell physiology Cells Contraceptive Agents Data Detection Development Dinoprostone Dose Endopeptidases 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 Role playing therapy 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

DESCRIPTION (provided by applicant): Ovulation is essential for successful reproduction. The ovulatory luteinizing hormone surge stimulates perieovulatory 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 perieovulatory 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 perieovulatory 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 expresses a unique subset of all EP receptors. Second, we will demonstrate that each EP receptor mediates a subset of all PGE2-stimulated preovulatory 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 preovulatory 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 preovulatory response to 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.

描述（由申请人提供）：排卵对于成功繁殖至关重要。排卵期黄体生成素激增刺激排卵期卵泡产生前列腺素 E2 (PGE2)，这是颗粒卵丘细胞扩张、卵泡破裂和卵母细胞释放所必需的。阻断 PGE2 的产生会导致排卵失败，这支持了对 PGE2 作为不孕症治疗方法的研究，以及作为潜在避孕药抑制 PGE2 合成/作用的研究。为了阐明 PGE2 刺激围排卵事件的机制，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中都具有独特的功能。首先，我们将确定哪些 EP 受体是由排卵卵泡内功能不同的颗粒细胞亚群表达的。在猴卵巢组织切片中使用激光捕获显微切割和免疫荧光 EP 检测，我们将识别存在于卵丘、基底壁和窦壁颗粒细胞以及位于卵泡顶端的颗粒细胞中的 EP 受体。这些亚群各自在排卵中发挥独特的作用，我们预计每个亚群表达所有 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 刺激的排卵前过程的一个子集。 PGE2 已知可刺激颗粒细胞黄体酮产生、卵丘扩张和卵泡破裂。将利用颗粒细胞培养物以及猕猴卵泡内注射来证明对每种 EP 受体具有特异性的激动剂会刺激部分但不是全部的 PGE2 介导的颗粒细胞功能。最后，我们将确定哪些 EP 受体可以在灵长类动物排卵前卵泡的颗粒细胞中转导 PGE2 信号。将用四种 EP 受体特异性激动剂处理颗粒细胞，并评估通过这些 G 蛋白偶联受体产生的细胞内信号。每个 EP 受体可能会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种 EP 受体介导灵长类卵泡内对 PGE2 的总排卵前反应的特定子集。这些数据将促进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，并可能最终导致不孕症新疗法和新避孕方法的开发。