Prostaglandin Receptors in Primate Ovulatory Follicles

灵长类动物排卵卵泡中的前列腺素受体

• 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
• 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

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.

排卵是成功繁殖的关键。排卵性促黄体激素激增刺激 排卵期卵泡产生前列腺素E2（PGE 2），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。阻止PGE 2的产生会导致排卵失败， 支持研究PGE 2作为不孕症的治疗以及PGE 2合成/作用的抑制 作为潜在的避孕药。为了阐明PGE 2刺激排卵期事件的机制， 该提案提出了一个总体假设，即四种PGE 2（EP）受体中的每一种都具有独特的功能， 排卵首先，我们将确定哪些EP受体由功能不同的亚群表达 排卵期卵泡内的颗粒细胞。使用激光捕获显微切割以及 在猴卵巢组织切片中进行免疫荧光EP检测，我们将鉴定EP受体 存在于卵丘、基底壁和窦壁颗粒细胞以及位于 毛囊顶端这些亚群在排卵过程中各自发挥着独特的作用，我们预计， 亚群表达所有EP受体的独特子集。第二，我们将证明每个EP 受体介导的一个子集的所有前列腺素E2刺激排卵期的过程。已知PGE 2会刺激 颗粒细胞孕酮产生、卵丘扩张和卵泡破裂。颗粒细胞培养 以及在猕猴中的滤泡内注射将用于证明对每种EP特异性的激动剂 受体刺激一些，但不是全部，这些必不可少的PGE 2介导的颗粒细胞功能。最后我们 将确定哪些EP受体可以阻断灵长类排卵期颗粒细胞中的PGE 2信号， 毛囊将用对四种EP受体中的每一种特异性的激动剂处理颗粒细胞，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受体将 可能以独特的细胞内反应响应激动剂刺激。拟议的研究可能会 表明每种EP受体介导对PGE 2的总排卵期反应的特定亚群 在灵长类卵泡中。这些数据将有助于有针对性的药理学方法， 刺激或抑制排卵过程的基本特征，并可能最终导致 不孕症的新疗法和新的避孕选择。