Regulation of Primordial Follicle Formation and Oocyte Survival

原始卵泡形成和卵母细胞存活的调节

• 批准号: 8879401
• 负责人: Melissa E Pepling
• 金额: $ 44.02万
• 依托单位: SYRACUSE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-17 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8879401
• 关键词: Affect; Age; Apoptosis; Apoptotic; BAX gene; BCL2 gene; Biology; Birth; Cell Count; Cell Death; Cell Differentiation process; Cells; Complex; Confocal Microscopy; Cyst; Data; Development; Disease; Embryonic Development; Equilibrium; Family member; Female; Female infertility; Foundations; Functional disorder; Future; Germ Cells; Goals; Gonadal structure; Grant; Infertility; Invertebrates; Knowledge; Laboratories; Life; Light; Longevity; MAP Kinase Gene; MCL1 gene; Malignant neoplasm of ovary; Menopause; Molecular; Molecular Target; Mus; Neonatal; Oocytes; Organ Culture Techniques; Ovarian; Ovarian Dysgerminoma; Ovarian Follicle; Ovary; Pathway interactions; Phase; Play; Population; Primordial Follicle; Process; Protein Family; Proteins; Receptor Protein-Tyrosine Kinases; Regulation; Reproduction; Research; Role; Signal Pathway; Signal Transduction; Small Interfering RNA; Structure of primordial sex cell; Techniques; Testing; Time; Vertebrates; Western Blotting; Woman; Work; base; design; fetal; genetic technology; granulosa cell; immunocytochemistry; improved; inhibitor/antagonist; mouse development; novel; premature; primary ovarian insufficiency; public health relevance; reproductive

 DESCRIPTION (provided by applicant): 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. Prior to birth, the oocytes develop in clusters called germline cysts, a conserved phase of oocyte development in both vertebrates and invertebrates. During late fetal and early neonatal development, mouse germ cell cysts break apart into single oocytes that become surrounded by pre-granulosa cells to form primordial follicles. During this 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 cyst breakdown, oocyte apoptosis and follicle assembly are currently unknown. The long-term goal is to understand molecular and cellular mechanisms that regulate cyst breakdown and programmed cell death to establish the primordial follicle pool in the mouse ovary. The objective of this proposal is to determine the role of two pathways, the KIT signaling pathway and the BCL2 apoptotic pathway in modulating cyst breakdown and oocyte numbers. The central hypothesis of the proposed research is that cyst breakdown and germ cell death are controlled by signaling through the receptor tyrosine kinase, KIT and by a balance of BCL2 pro- and anti-apoptotic proteins. Recent work from our laboratory suggests KIT signaling may play an important role. In addition, we provide preliminary data demonstrating that BCL2 family proteins are also important for the regulation of cyst breakdown and associated programmed cell death. This proposal explores the molecular and cellular aspects of KIT signaling and programmed cell death regulators in cyst breakdown and oocyte survival. The specific aims of this research are to: 1) elucidate the molecular and cellular mechanisms of KIT signaling in cyst breakdown and associated oocyte loss; and 2) identify BCL2 family members involved in regulating oocyte survival. These goals will be achieved through techniques including immunocytochemistry, confocal microscopy, Western blotting, ovary organ culture, pharmacological inhibitors, siRNA technology and genetics. Research proposed in the current application is significant because it will enhance our current knowledge by elucidating the mechanisms by which cyst breakdown and associated oocyte loss are regulated. 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.

 描述（由适用提供）：尽管对物种的延续很重要，但原始生殖细胞分化为功能性卵母细胞的分化知之甚少。原始生殖细胞在小鼠的胚胎发育过程中开始分化为卵母细胞。出生前，卵母细胞在称为种系囊肿的簇中发育，这是脊椎动物和无脊椎动物中卵母细胞发育的保守阶段。在胎儿晚期和新生儿发育期间，小鼠生殖细胞囊肿分解成单个卵母细胞，这些卵母细胞被颗粒前细胞周围形成以形成原始锻造。在此囊肿分解过程中，每个囊肿中的一个细胞的子集死亡，只有三分之一的初始数量存活以形成原始的伪造。目前尚不清楚控制囊肿分解，圆凋亡和叶叶组件的机制。长期目标是了解调节囊肿分解和程序性细胞死亡以在小鼠卵巢中建立原始叶池的分子和细胞机制。该提案的目的是确定两种途径的作用，即套件信号通路和BCL2凋亡途径在调节囊肿分解和卵母细胞数中的作用。拟议研究的中心假设是，通过通过受体酪氨酸激酶，试剂盒和Bcl2 pro和抗凋亡蛋白的平衡来控制囊肿分解和生殖细胞死亡。我们实验室的最新工作表明，套件信号传导可能起着重要作用。此外，我们还提供了初步数据，表明Bcl2家族蛋白对于调节囊肿分解和相关的编程细胞死亡也很重要。该提案探讨了在囊肿分解和卵母细胞存活中，试剂盒信号传导和程序性细胞死亡调节剂的分子和细胞方面。这项研究的具体目的是：1）在囊肿分解和相关的卵母细胞损失中阐明试剂盒信号传导的分子和细胞机制； 2）确定参与调节卵母细胞存活的BCL2家族成员。这些目标将通过包括免疫细胞化学，共聚焦显微镜，蛋白质印迹，卵巢器官培养，药物抑制剂，siRNA技术和遗传学等技术实现。当前应用中提出的研究非常重要，因为它将通过阐明囊肿分解和相关卵母细胞损失的机制来增强我们当前的知识。在这笔赠款中获得的结果将有助于改善卵巢生物学的研究工作和治疗导致女性不孕症（例如原发性卵巢症）的疾病的治疗。