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 尽管对物种的延续很重要，但原始的分化 生殖细胞成功能性卵母细胞的理解很少。原始生殖细胞开始 在小鼠的胚胎发育过程中分化为卵母细胞。卵母细胞在 簇称为种系囊肿，这是两个脊椎动物中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的预言并在副烯中逮捕 阶段。然后，他们发生原始卵泡形成，在此期间生殖细胞囊肿分解 进入单个卵母细胞（囊肿分解）和颗粒细胞围绕单个卵母细胞迁移到 形成原始卵泡。在囊肿分解过程中，每个囊肿死亡的一个细胞子集 只有三分之一的初始卵母细胞存活以形成原始卵泡。这 控制减数分裂进展，囊肿分解，颗粒细胞募集和 卵母细胞的生存尚不清楚。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。这 该建议的目的是了解套件信号通路在调节中的作用 减数分裂预言I和原始卵泡形成。提议的中心假设 研究是从受体酪氨酸激酶Kit发出的信号传导促进原始卵泡 通过调节 下游目标。我们实验室的最新工作表明，套件信号可能会发挥 重要角色。该建议探讨了套件信号传导的分子和细胞方面 建立原始卵泡池。这项研究的具体目的是：1）阐明 试剂盒信号转导在原始卵泡形成和卵母细胞进展中的作用 减数分裂预言i； 2）确定在卵母细胞中重要的套件下游目标 发展。这些目标将通过包括 免疫组织化学，共聚焦显微镜，卵巢器官培养，实时PCR和单细胞 RNA测序分析。当前应用中提出的研究很重要，因为它 将通过阐明建立重要的机制来增强我们当前的知识 原始卵泡池。在本赠款中获得的结果将有助于改善研究工作 卵巢生物学以及在造成女性不育症（例如原发性卵巢）的疾病的治疗中 不足。