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.

项目描述（由申请人提供）：本申请是关于卵母细胞成熟和受精过程中的信号转导的延续。它解决了长期悬而未决的问题，促黄体生成素（LH）如何导致哺乳动物卵泡封闭的卵母细胞减数分裂恢复。最近的研究结果表明，卵泡体细胞之间的缝隙连接在LH的作用下迅速关闭，这构成了本项目3个目标的基础：1）研究LH诱导的卵泡间隙连接闭合是否由TACE蛋白酶介导的EGF受体活化引起，2）研究卵泡细胞中磷酸化介导的Cx43缝隙连接的关闭是否是响应LH恢复减数分裂所必需的，3）探讨缝隙连接关闭在LH刺激下卵母细胞cAMP和cGMP调节中的作用。将通过显微注射含有荧光示踪剂的活卵泡封闭的卵母细胞，并通过双光子显微镜和光漂白后的荧光恢复进行分析，研究间隙连接通透性。这些研究将通过对连接蛋白43在特定调控位点上的磷酸化的研究来补充。将使用新开发的光学探针和共聚焦显微镜监测卵泡封闭的卵母细胞中cAMP和cGMP的浓度。这些活组织显微镜检查方法是一个重大进步，因为它们避免了为了研究其功能而破坏卵泡调节环境的必要性。将使用转基因小鼠，与这些显微镜方法相结合，以检验EGF受体配体的蛋白水解加工和连接蛋白43在已鉴定丝氨酸上的磷酸化对于从卵泡体细胞到卵母细胞的信号转导是必不可少的这一假设。这项拟议中的研究将推进对一个关键生物过程的认识，并为未来的临床发展奠定基础，特别是在治疗不孕症方面。卵母细胞体外成熟是人类体外受精方法的一个新兴组成部分，对控制卵母细胞减数分裂的信号机制的理解将促进这种进步。公共卫生相关性：这项拟议中的研究涉及激素信号导致储存在卵巢中的哺乳动物卵母细胞发育到可受精阶段的机制。这项研究将推进对一个关键生物过程的认识，并为未来的临床发展奠定基础，特别是在治疗不孕症方面。