Follicular morphogenesis during perinatal development

围产期发育过程中的卵泡形态发生

基本信息

批准号：
8091316
负责人：
SHYAMAL K. ROY
金额：
$ 25.26万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-07-01 至 2012-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8091316
关键词：
Address Affect Antibodies Attenuated Cells Communication Complement Factor B Complementary DNA Contraceptive methods Cues Defect Development Developmental Process ESR1 gene ESR2 gene Endocrine Estradiol Estrogen Receptors Event Exposure to Fertility Fertilization Fetal Development Funding G-Protein-Coupled Receptors Goals Growth Differentiation Factor 9 Hamsters Human In Vitro Infertility Knowledge Laboratories Lead Life Modeling Molecular Morphogenesis Mutation Neonatal Oocytes Outcome Ovarian Ovary Ovulation Perinatal Perinatal Exposure Physiological Play Premature Ovarian Failure Primordial Follicle Proteins Protocols documentation RNA Interference Regulation Research Role Signal Transduction Small Interfering RNA Somatic Cell Sterility System Testing Therapeutic Transforming Growth Factors Transgenes Woman base bone morphogenetic protein receptor type II egg fetal folliculogenesis granulosa cell improved in vivo insight intraovarian novel therapeutics paracrine postnatal prenatal receptor receptor expression reproductive tool

项目摘要

DESCRIPTION (provided by applicant): Formation and development of primordial follicles are obligatory events for successful folliculogenesis and fertility. Defects in these steps lead to ovarian dysgenesis or premature ovarian failure and ultimately infertility. Our long-term goal is to elucidate the regulatory mechanisms controlling the formation and development of the primordial follicles as a necessary prerequisite to the development of improved therapeutic management of human infertility and contraception. Although evidence suggests that growth differentiation factor 9 (GDF9) or E2 stimulates preantral follicle development, virtually nothing is known about their role in primordial folliculogenesis or the mechanisms therein. This renewal application is focused on filling this knowledge gap. During the last funded period, we have shown that inactivation of FSH during fetal development disrupts primordial folliculogenesis, which correlates with low expression of GDF9 and estrogen receptors (ESR). Further, FSH treatment upregulates GDF9 and ESR expression. Our preliminary results indicate that (1) suppression of endogenous GDF9 expression in vitro retards primordial folliculogenesis, (2) E2 stimulates primordial follicle formation in vitro, (3) fetal exposure to an FSHantiserum blocks the expression of the GDF9 receptors, such as bone morphogenetic protein receptor II [BMPRII] and transforming growth factor B receptor type I [TBRI], and blocks the expression of ESR1 and ESR2, and (4) siRNA knockdown of a G-protein coupled receptor 30 (GPR30), a transmembrane ESR in vitro attenuates the formation of primordial follicles. Based on these observations we hypothesize that GDF9 and E2 promote the differentiation of somatic cells into granulosa cells leading to the formation of primordial follicles by mechanisms that involve a concerted action of the GDF9 receptors (BMPRII and TBRI), and the ESR (ESR1, ESR2 and GPR30). We further postulate that FSH regulates the action of GDF9 and E2 by modulating the expression of their receptors. The hypothesis will be tested with the following specific aims: 1. To reveal the physiological importance of GDF9 expression in the differentiation of somatic cells into pregranulosa cells leading to primordial follicle formation. We will determine the need for intraovarian GDF9, and its mechanisms of action. 2. To examine the physiological importance of the ESR in the differentiation of somatic cells into pregranulosa cells leading to primordial follicle formation. We will determine the need for the classic ESR and GPR30 with respect to GDF9 action. We will use fetal and neonatal hamster ovaries in vivo and in vitro (primordial follicles do not appear until 8th day of life), RNAi, antisense oligodeoxynucleotides, pharmacological, morphological and molecular approaches to address the specific aims. The successful completion of this project should advance our understanding of the mechanisms involved in the morphogenesis of primordial follicles. Lay summary: The primordial follicle stock represents a nonrenewable follicular reserve for the entire reproductive life of the mammalian species, including the human, and determines the fertility and fecundity of the species. The purpose of the proposed research is to elucidate the regulatory mechanisms controlling the formation and development of the primordial follicles. The outcomes will have a significant impact on the development of improved or novel therapeutic management of human infertility and contraception.

描述（由申请人提供）：原始卵泡的形成和发育是成功卵泡发生和生育的必要事件。这些步骤的缺陷会导致卵巢发育不全或卵巢早衰，并最终导致不孕。我们的长期目标是阐明控制原始卵泡形成和发育的调节机制，这是改进人类不孕和避孕治疗管理的必要先决条件。尽管有证据表明生长分化因子 9 (GDF9) 或 E2 刺激窦前卵泡发育，但实际上对其在原始卵泡发生中的作用或其中的机制一无所知。此更新应用程序的重点是填补这一知识空白。在上一个资助期间，我们发现胎儿发育过程中 FSH 失活会破坏原始卵泡发生，这与 GDF9 和雌激素受体 (ESR) 的低表达相关。此外，FSH 治疗上调 GDF9 和 ESR 表达。我们的初步结果表明，(1) 体外抑制内源性 GDF9 表达会延迟原始卵泡发生，(2) E2 刺激体外原始卵泡形成，(3) 胎儿暴露于 FSH 抗血清会阻断 GDF9 受体的表达，例如骨形态发生蛋白受体 II [BMPRII] 和转化生长因子 B 受体 I 型 [TBRI]，并阻断 ESR1 和 ESR2 的表达，以及(4) siRNA 敲低 G 蛋白偶联受体 30 (GPR30)（一种体外跨膜 ESR）会减弱原始卵泡的形成。基于这些观察，我们假设 GDF9 和 E2 通过涉及 GDF9 受体（BMPRII 和 TBRI）和 ESR（ESR1、ESR2）协同作用的机制促进体细胞分化为颗粒细胞，从而导致原始卵泡的形成。和 GPR30）。我们进一步假设 FSH 通过调节 GDF9 和 E2 受体的表达来调节 GDF9 和 E2 的作用。该假设将通过以下具体目标进行检验： 1. 揭示 GDF9 表达在体细胞分化为前颗粒细胞并导致原始卵泡形成中的生理重要性。我们将确定卵巢内 GDF9 的需求及其作用机制。 2. 检查ESR在体细胞分化为前颗粒细胞并导致原始卵泡形成中的生理重要性。我们将确定 GDF9 动作是否需要经典 ESR 和 GPR30。我们将使用胎儿和新生儿仓鼠卵巢的体内和体外（原始卵泡直到生命第 8 天才出现）、RNAi、反义寡脱氧核苷酸、药理学、形态学和分子方法来解决特定目标。该项目的成功完成应增进我们对原始卵泡形态发生机制的理解。摘要：原始卵泡储备代表了哺乳动物（包括人类）整个生殖生命的不可再生的卵泡储备，并决定了该物种的生育力和繁殖力。本研究的目的是阐明控制原始卵泡形成和发育的调节机制。研究结果将对人类不孕症和避孕的改进或新型治疗管理的发展产生重大影响。