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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要/摘要尽管原始物种的分化对于物种的延续很重要，人们对于生殖细胞如何转化为功能性卵母细胞知之甚少。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。卵母细胞发育于称为种系囊肿的簇，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 并在双线期停滞阶段。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（囊肿破裂），颗粒细胞在单个卵母细胞周围迁移形成原始卵泡。在囊肿破裂的过程中，每个囊肿中的一部分细胞死亡只有最初数量的三分之一的卵母细胞能够存活下来形成原始卵泡。这控制减数分裂进展、囊肿破裂、颗粒细胞募集和卵母细胞的存活率尚不清楚。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。这该提案的目的是了解 KIT 信号通路在调节中的作用减数分裂前期 I 和原始卵泡的形成。所提出的中心假设研究表明，受体酪氨酸激酶 KIT 的信号传导可促进原始卵泡通过调节卵母细胞的形成和进展到前期I的双线期阶段下游目标。我们实验室最近的工作表明 KIT 信号传导可能发挥着重要作用重要作用。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的作用减数分裂前期I； 2) 确定 KIT 下游在卵母细胞生成过程中重要的靶点发展。这些目标将通过以下技术实现：免疫组织化学、共聚焦显微镜、卵巢器官培养、实时 PCR 和单细胞 RNA测序分析。当前申请中提出的研究很重要，因为它将通过阐明建立原始卵泡池。这笔赠款中获得的成果将有助于改善以下方面的研究工作卵巢生物学和治疗导致女性不孕的疾病，例如原发性卵巢不足。