CIPAR1, A Novel Modulator of Progesterone Accumulation in Periovulatory Follicle

CIPAR1，排卵周围卵泡黄体酮积累的新型调节剂

• 批准号: 8609430
• 负责人: MISUNG JO
• 金额: $ 16.83万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8609430
• 关键词: Affect; Antibodies; Apoptosis; Castration; Cell Culture Techniques; Cell Differentiation process; Cell physiology; Cells; Collaborations; Cytoprotection; Data; Development; Enzymes; Experimental Models; Female; Fertility; Future; Gene Proteins; Genes; Goals; Gonadotropins; Hormones; Human; Immunoprecipitation; In Vitro; Instruction; Link; Location; Luteinization; Macaca; Mediating; Mediator of activation protein; Molecular; Monkeys; Ovary; Ovulation; Physiological; Play; Primates; Production; Progesterone; Prostaglandins; Prostate; Prostatic; Proteins; Proteomics; Rattus; Reproduction; Rodent; Role; Steroid biosynthesis; Testing; Tissues; Woman; base; corpus luteum; critical period; granulosa cell; hormone regulation; human tissue; improved; in vitro Model; in vivo; novel; research study; response

PROJECT SUMMARY (See instructions): In response to the preovulatory LH surge, the preovulatory follicle rapidly increases progesterone production, which is essential for ovulation and/or corpus luteum formation. Although this rise in progesterone level has been mainly linked to a few LH-induced genes involved in steroidogenesis (e.g., StAR, Cypi 1 a l , and HSD3b), but there are many gaps in our understanding of periovulatory accumulation of progesterone. In this proposal, we put forward a novel protein, CIPAR1 (castration-induced prostatic apoptosis-related protein 1), as a vital mediator of progesterone accumulation in the periovulatory follicle. We have recently demonstrated the LH surge increases the expression of Ciparl in periovulatory follicles of rodent ovaries. More importantly, our preliminary study using granulosa cell cultures showed that knockdown of Ciparl expression resulted in significant reduction of progesterone levels. Based on these novel findings, we hypothesize that induction of CIPAR1 by the LH surge is crucial for progesterone accumulation in periovulatory follicles, thus ovulation and luteinization. In specific Aim#1, we will demonstrate that the alteration of Ciparl expression by silencing or over-expression affects progesterone production in rat periovulatory follicles using both in vivo and in vitro models. As the first approach to pinpoint the functional contribution of C1PAR1 in periovulatory follicles, we will identify the genes and proteins that are differentially expressed when Ciparl expression is silenced or over-expressed in periovulatory follicles. Because littie is known about cellular function of CIPAR1, Specific Aim #2 focuses on first determining the cellular location of CIPAR1 in periovulatory granulosa cells using confocal co-localization and/or subcellular fractionated protein analyses and then identifying CIPAR1 interacting proteins in periovulatory granulosa cells using immunoprecipitation, followed by a proteomic approach. Other than our preliminary data detecting Ciparl expression in human granulosa cells, nothing is known about CIPAR1 in human tissues. In collaboration with Drs. Brannstrum and Duffy, specific Aim #3 will determine whether Ciparl expression is hormonally regulated during the periovulatory period and critical for LH-induced progesterone production in humans and/or macaques. Data obtain from the proposed studies will unravel the role of C1PAR1 as a novel and critical mediator of progesterone accumulation in periovulatory follicles of primates as well as rodents.

项目总结（见说明）： 作为对排卵前LH峰的反应，排卵前卵泡迅速增加孕酮的产生，这对排卵和/或黄体形成是必需的。虽然孕酮水平的这种升高主要与一些LH诱导的参与类固醇生成的基因有关（例如，星星、Cypi 1a 1和HSD 3b），但在我们对排卵期孕酮积累的理解中存在许多空白。 在这个建议中，我们提出了一种新的蛋白质，CIPAR 1（去势诱导的前列腺增生相关蛋白1），作为一个重要的调解孕激素积累在排卵期卵泡。我们最近已经证明LH峰增加啮齿动物卵巢排卵期卵泡中Ciparl的表达。更重要的是，我们使用颗粒细胞培养物的初步研究表明，Ciparl表达的敲低导致孕酮水平的显着降低。基于这些新的发现，我们假设CIPAR 1的LH峰诱导是至关重要的孕激素积累在排卵期卵泡，从而排卵和黄体化。在具体的目标#1中，我们将使用体内和体外模型证明通过沉默或过表达改变Ciparl表达影响大鼠排卵期卵泡中的孕酮产生。作为确定C1 PAR 1在排卵期卵泡中的功能贡献的第一种方法，我们将鉴定当Ciparl表达在排卵期卵泡中沉默或过表达时差异表达的基因和蛋白质。由于对CIPAR 1的细胞功能知之甚少，具体目标#2侧重于首先确定CIPAR 1的细胞位置。 使用共聚焦共定位和/或亚细胞分级蛋白分析，然后使用免疫沉淀法，随后通过蛋白质组学方法鉴定排卵期颗粒细胞中的CIPAR 1相互作用蛋白。除了我们检测到Ciparl在人类颗粒细胞中表达的初步数据外，对CIPAR 1在人类组织中的表达一无所知。在Brannstrum和Duffy博士的合作下，具体目标#3将确定Ciparl表达是否与细胞增殖有关。 在排卵期调节，对LH诱导的人和/或猕猴孕酮产生至关重要。从拟议的研究中获得的数据将揭示C1 PAR 1作为灵长类和啮齿类动物排卵期卵泡中孕酮积累的新的关键介质的作用。