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.

项目摘要（参见说明）： 为了响应排卵前 LH 激增，排卵前卵泡迅速增加黄体酮的产生，这对于排卵和/或黄体形成至关重要。尽管黄体酮水平的升高主要与一些涉及类固醇生成的 LH 诱导基因（例如 StAR、Cypi 1 a1 和 HSD3b）有关，但我们对排卵期黄体酮积累的理解存在许多差距。 在这项提案中，我们提出了一种新的蛋白质 CIPAR1（去势诱导的前列腺细胞凋亡相关蛋白 1），作为排卵期卵泡中黄体酮积累的重要介质。我们最近证明 LH 激增会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进行的初步研究表明，Ciparl 表达的敲低会导致黄体酮水平显着降低。基于这些新发现，我们假设 LH 激增诱导 CIPAR1 对于排卵周围卵泡中黄体酮的积累，从而促进排卵和黄体化至关重要。在具体目标#1中，我们将使用体内和体外模型证明通过沉默或过度表达来改变 Ciparl 表达会影响大鼠排卵期卵泡中孕酮的产生。作为确定 C1PAR1 在排卵期卵泡中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度表达时差异表达的基因和蛋白质。因为我们知道 CIPAR1 的细胞功能，所以具体目标 #2 侧重于首先确定 CIPAR1 的细胞位置 使用共聚焦共定位和/或亚细胞分级蛋白分析分析排卵周围颗粒细胞中的 CIPAR1，然后使用免疫沉淀法鉴定排卵周围颗粒细胞中的 CIPAR1 相互作用蛋白，然后采用蛋白质组学方法。除了我们检测人类颗粒细胞中 Ciparl 表达的初步数据外，我们对人类组织中的 CIPAR1 一无所知。与博士合作。 Brannstrum 和 Duffy，具体目标 #3 将确定 Ciparl 表达是否是激素性的 在排卵期期间受到调节，对于人类和/或猕猴中 LH 诱导的黄体酮产生至关重要。从拟议研究中获得的数据将揭示 C1PAR1 作为灵长类动物和啮齿类动物排卵期卵泡中黄体酮积累的新型关键介质的作用。