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）：原始卵泡的形成和开发是成功的卵泡发生和生育能力的强制性事件。这些步骤中的缺陷导致卵巢发病障碍或卵巢过早的失败，并最终导致不孕症。我们的长期目标是阐明控制原始卵泡形成和发展的调节机制，这是发展人类不育和避孕的治疗性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刺激了毛囊的发育，但实际上对它们在原始卵泡发生或其中的机制中的作用几乎一无所知。该更新应用程序的重点是填补这一知识差距。在最后的资助时期，我们表明，胎儿发育过程中FSH的失活破坏了原始卵泡发生，这与GDF9和雌激素受体（ESR）的低表达相关。此外，FSH处理上调GDF9和ESR表达。 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]，并阻止ESR1和ESR2的表达，以及（4）G蛋白耦合受体30（GPR30）的siRNA敲低，这是一种体外跨膜ESR的siRNA耦合受体30（GPR30），可减弱原始卵泡的形成。基于这些观察结果，我们假设GDF9和E2促进了体细胞向颗粒细胞的分化，从而通过机制形成了原始卵泡的形成，该机制涉及GDF9受体（BMPRII和TBRI）的一致作用，以及ESR1，ESR1，ESR2和GPR30。我们进一步假设FSH通过调节其受体的表达来调节GDF9和E2的作用。该假设将以以下特定目的进行检验：1。揭示GDF9表达在细胞分化为粒细胞中的生理重要性，从而导致原始卵泡形成。我们将确定对摩var内GDF9及其作用机理的需求。 2。检查ESR在细胞分化为粒细胞中的生理重要性，从而导致原始卵泡形成。我们将确定对GDF9动作的经典ESR和GPR30的需求。我们将在体内和体外使用胎儿和新生儿仓鼠卵巢（原始卵泡直到生命的第8天才出现），RNAi，反义寡脱氧核苷酸，药理，形态学和分子方法可解决特定目标。该项目的成功完成应提高我们对原始卵泡形态发生的机制的理解。摘要摘要：原始卵泡库存代表了包括人类在内的哺乳动物物种的整个生殖寿命的不可再生卵泡储备，并确定了该物种的生育能力和生育能力。拟议研究的目的是阐明控制原始卵泡形成和发展的调节机制。结果将对人类不育症和避孕的改善或新颖的治疗管理的发展产生重大影响。