Signal Transduction at Fertilization

受精时的信号转导

• 批准号: 8134307
• 负责人: LAURINDA A. JAFFE
• 金额: $ 29.7万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-03-01 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8134307
• 关键词: Accounting; Address; Biological Process; Cell membrane; Cells; Clinical; Communication; Complement; Confocal Microscopy; Connexin 43; Connexins; Cyclic AMP; Cyclic GMP; Cyclic Nucleotides; Development; Diffusion; Environment; Epidermal Growth Factor Receptor; Event; Fertilization; Fertilization in Vitro; Fluorescence Recovery After Photobleaching; Future; G-Protein-Coupled Receptors; GTP-Binding Proteins; Gap Junctions; Health; Hormonal; Human; Imaging Techniques; In Vitro; Infertility; Investigation; Knock-in Mouse; Knowledge; Life; Ligands; Luteinizing Hormone; Measurement; Measures; Mediating; Meiosis; Methods; Microinjections; Microscopic; Microscopy; Modification; Monitor; Mus; Oocytes; Optical Methods; Optics; Ovary; Peptide Hydrolases; Permeability; Phosphorylation; Photons; Property; Prophase; Proteolytic Processing; Receptor Activation; Research; Role; Serine; Signal Transduction; Site; Somatic Cell; Staging; Testing; Tissues; Tracer; base; computerized data processing; granulosa cell; mutant; oocyte maturation; prevent; research study; response

DESCRIPTION (provided by applicant): This application is for renewal of a project concerned with signal transduction during oocyte maturation and fertilization. It addresses the long-standing unanswered question of how luteinizing hormone (LH) causes meiotic resumption in mammalian follicle- enclosed oocytes. Recent findings, which establish that gap junctions between the somatic cells of the follicle close rapidly in response to LH, form the basis for the 3 aims of this project: 1) to investigate if LH-induced gap junction closure in the follicle is caused by TACE protease-mediated activation of the EGF receptor, 2) to investigate whether phosphorylation-mediated closure of Cx43 gap junctions in the follicle cells is required for resumption of meiosis in response to LH, and 3) to investigate the role of gap junction closure in regulating cAMP and cGMP in the oocyte in response to LH. Gap junction permeability will be investigated by microinjection of live follicle-enclosed oocytes with fluorescent tracers, and analysis by 2-photon microscopy and fluorescence recovery after photobleaching. These studies will be complemented by investigations of the phosphorylation of connexin 43 on specific regulatory sites. Concentrations of cAMP and cGMP will be monitored in follicle-enclosed oocytes using newly developed optical probes and confocal microscopy. These methods for live tissue microscopy are a major advance, because they avoid the necessity of disrupting the regulatory environment of the follicle in order to investigate its function. Genetically modified mice will be used, in combination with these microscopic approaches, in order to test the hypothesis that proteolytic processing of EGF receptor ligands and phosphorylation of connexin 43 on identified serines are essential for transduction of signals from the somatic cells of the follicle to the oocyte. The proposed research will advance knowledge of a crucial biological process, and establish a basis for future clinical developments, especially in the treatment of infertility. In vitro oocyte maturation is an emerging component of methods for human in vitro fertilization, and understanding of signaling mechanisms that control oocyte meiosis will facilitate such advances. PUBLIC HEALTH RELEVANCE: The proposed research concerns the mechanisms by which hormonal signals cause mammalian oocytes, stored in the ovary, to develop to the fertilizable stage. This research will advance knowledge of a crucial biological process, and establish a basis for future clinical developments, especially in the treatment of infertility.

描述（由申请人提供）：此申请是用于续签卵母细胞成熟和受精过程中有关信号转导的项目。它解决了长期以来的未解决的问题，即黄体生成激素（LH）如何导致哺乳动物卵泡封闭的卵母细胞中的减数分裂恢复。 Recent findings, which establish that gap junctions between the somatic cells of the follicle close rapidly in response to LH, form the basis for the 3 aims of this project: 1) to investigate if LH-induced gap junction closure in the follicle is caused by TACE protease-mediated activation of the EGF receptor, 2) to investigate whether phosphorylation-mediated closure of Cx43 gap junctions in the follicle cells is required for响应LH的减数分裂以及3）研究间隙连接闭合在调节CAMP和CGMP中响应LH的作用。间隙连接渗透性将通过与荧光示踪剂的活卵泡封闭的卵母细胞的显微注射进行研究，并通过2光子显微镜和荧光在光漂白后进行分析。这些研究将通过对特定调节部位上连接蛋白43的磷酸化的研究来补充。 CAMP和CGMP的浓度将在卵泡封闭的卵母细胞中使用新开发的光学探针和共聚焦显微镜进行监测。这些用于活组织显微镜的方法是一项重大进展，因为它们避免了破坏卵泡的调节环境以研究其功能的必要性。结合这些微观方法，将使用遗传改性的小鼠，以检验以下假设：EGF受体配体的蛋白水解处理以及鉴定出的连接蛋白43的磷酸化对于从卵泡细胞到卵母细胞的躯体细胞的信号转导至关重要。拟议的研究将提高对关键生物学过程的了解，并为未来的临床发展建立基础，尤其是在治疗不孕症方面。体外卵母细胞的成熟是人类体外受精方法的新兴成分，了解控制卵母细胞减数分裂的信号传导机制将促进此类进步。公共卫生相关性：拟议的研究涉及激素信号导致卵巢储存的哺乳动物卵母细胞的机制，以发展为可肥沃的阶段。这项研究将促进对关键生物学过程的了解，并为未来的临床发展建立基础，尤其是在不孕症治疗方面。