Mechanisms of KIT signaling in the regulation of primordial follicle formation

项目摘要/摘要 尽管它对物种的延续很重要，但原始物种的分化 生殖细胞转化为有功能的卵母细胞还知之甚少。原始生殖细胞开始 在小鼠胚胎发育过程中分化为卵母细胞。卵母细胞在体内发育 在这两种脊椎动物中，卵子发育的一个保守阶段称为生殖系囊泡。 和无脊椎动物。卵母细胞经过减数分裂前期I和双线期停滞 舞台。然后他们经历原始卵泡的形成，在这个过程中生殖细胞囊分裂 进入单个卵母细胞(囊破裂)，颗粒细胞在单个卵母细胞周围迁移到 形成原始毛囊。在包囊破裂的过程中，每个包囊中的一部分细胞死亡 只有初始数量的三分之一的卵母细胞存活下来，形成原始卵泡。这个 控制减数分裂进展、囊泡破裂、颗粒细胞募集和 卵母细胞的存活还不是很清楚。我们的长期目标是了解分子和 在小鼠卵巢中建立原始卵泡池的细胞机制。这个 这项建议的目的是了解KIT信号通路在调节 减数分裂前期I和原始卵泡的形成。建议中的中心假设 研究表明，受体酪氨酸激酶KIT发出的信号可以促进原始卵泡 调控卵母细胞向双线期的形成和发育 下游目标。我们实验室最近的研究表明，试剂盒信号可能在 重要的角色。这项提案探讨了KIT信号转导的分子和细胞方面 建立原始卵泡池。本研究的具体目的是：1)阐明 KIT信号转导通路在原始卵泡形成和卵母细胞发育中的作用 减数分裂前期I；和2)确定KIT下游在卵母细胞过程中重要的靶点 发展。这些目标将通过包括以下技术实现 免疫组织化学、共聚焦显微镜、卵巢器官培养、实时定量聚合酶链式反应和单细胞 RNA测序分析。在当前应用中提出的研究具有重要意义，因为它 将通过阐明建立 原始毛囊池。这项资助所取得的成果将有助于改善 卵巢生物学及其在原发卵巢等女性不孕症治疗中的应用 不够用。

PROJECT SUMMARY/ABSTRACT Despite its importance to the continuation of species, the differentiation of primordial germ cells into functional oocytes is poorly understood. Primordial germ cells begin to differentiate into oocytes during embryonic development in the mouse. The oocytes develop in clusters called germline cysts, a conserved phase of oocyte development in both vertebrates and invertebrates. Oocytes progress through prophase I of meiosis and arrest at the diplotene stage. They then undergo primordial follicle formation during which germ cell cysts break apart into single oocytes (cyst breakdown) and granulosa cells migrate around individual oocytes to form primordial follicles. During the process of cyst breakdown, a subset of cells in each cyst die with only a third of the initial number of oocytes surviving to form primordial follicles. The mechanisms that control meiotic progression, cyst breakdown, granulosa cell recruitment and oocyte survival are not well understood. Our long-term goal is to understand molecular and cellular mechanisms used to establish the primordial follicle pool in the mouse ovary. The objective of this proposal is to understand the role of the KIT signaling pathway in regulating meiotic prophase I and primordial follicle formation. The central hypothesis of the proposed research is that signaling from the receptor tyrosine kinase, KIT, promotes primordial follicle formation and oocyte progression to the diplotene stage of prophase I through the regulation of downstream targets. Recent work from our laboratory suggests KIT signaling may play an important role. This proposal explores the molecular and cellular aspects of KIT signaling in establishing the pool of primordial follicles. The specific aims of this research are to: 1) elucidate the role of KIT signal transduction in primordial follicle formation and oocyte progression through meiotic prophase I; and 2) identify targets downstream of KIT that are important during oocyte development. These goals will be achieved through techniques including immunohistochemistry, confocal microscopy, ovary organ culture, real time PCR and single cell RNA sequencing analysis. Research proposed in the current application is significant because it will enhance our current knowledge by elucidating the mechanisms important to establish the primordial follicle pool. Results obtained in this grant will help improve research efforts in ovarian biology and in treatment of conditions causing female infertility such as primary ovarian insufficiency.