Mechanisms of KIT signaling in the regulation of primordial follicle formation

KIT信号调节原始卵泡形成的机制

• 批准号: 10680534
• 负责人: Melissa E Pepling
• 金额: $ 7.45万
• 依托单位: SYRACUSE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-09 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10680534
• 关键词: Affect; Biology; Birth; Cells; Confocal Microscopy; Cyst; Cytoplasm; Data; Development; Diplotene Stage; Embryo; Embryonic Development; Female; Female infertility; Foundations; Future; Genital; Genitalia; Germ Cells; Goals; Grant; Immunohistochemistry; Individual; Invertebrates; Knowledge; Laboratories; Life; Lighting; Longevity; Malignant neoplasm of ovary; Meiosis; Meiotic Prophase I; Menopause; Mitotic; Molecular; Mus; Oocytes; Oogonia; Organ Culture Techniques; Outcome; Ovarian; Ovarian Dysgerminoma; Ovary; Phase; Play; Population; Primordial Follicle; Process; Quality Control; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Regulation; Reproduction; Research; Role; Signal Pathway; Signal Transduction; Structure of primordial sex cell; Techniques; Testing; Time; Vertebrates; Work; cell motility; design; female fertility; granulosa cell; idiopathic infertility; improved; infertility treatment; intercellular connection; novel; ovarian dysfunction; ovarian reserve; premature; primary ovarian insufficiency; recruit; reproductive; reproductive system disorder; single-cell RNA sequencing

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.

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 生殖细胞转化为功能性卵母细胞的机制知之甚少。原始生殖细胞开始 在小鼠胚胎发育过程中分化成卵母细胞。卵母细胞发育在 称为种系囊肿的簇，是两种脊椎动物卵母细胞发育的保守阶段 和无脊椎动物。卵母细胞通过减数分裂前期I并停滞在双线期 阶段然后，它们经历原始卵泡形成，在此期间，生殖细胞囊肿破裂 进入单个卵母细胞（包囊破裂），颗粒细胞围绕单个卵母细胞迁移， 形成原始卵泡。在包囊破裂的过程中，每个包囊中的一个细胞亚群死亡 只有三分之一的卵母细胞存活下来形成原始卵泡。的 控制减数分裂进程、包囊破裂、颗粒细胞募集和 卵母细胞的存活还不清楚。我们的长期目标是了解分子和 用于在小鼠卵巢中建立原始卵泡池的细胞机制。的 本提案的目的是了解KIT信号通路在调节 减数分裂前期I和原始卵泡形成。提出的中心假设 研究表明，来自受体酪氨酸激酶KIT的信号促进原始卵泡发育， 形成和卵母细胞进展到双线期阶段的前期I通过调节 下游目标我们实验室最近的工作表明，KIT信号可能在 重要的作用该提案探讨了KIT信号传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信号转导在原始卵泡形成和卵母细胞发育中作用 减数分裂前期I;和2）鉴定在卵母细胞发育过程中重要的KIT下游靶标 发展这些目标将通过以下技术来实现， 免疫组化、共聚焦显微镜、卵巢器官培养、真实的时间PCR和单细胞 RNA测序分析。本申请中提出的研究是重要的，因为它 将通过阐明重要的机制来加强我们目前的知识， 原始卵泡池在这项补助金中获得的成果将有助于改善研究工作， 卵巢生物学和治疗引起女性不孕症的病症， 不足