SIGNAL TRANSDUCTION AT FERTILIZATION

受精时的信号转导

• 批准号: 3312873
• 负责人: LAURINDA A. JAFFE
• 金额: $ 12.65万
• 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 1996-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3312873
• 关键词: Asteroidea; DNA replication; G protein; Rana; Xenopus oocyte; biological signal transduction; calcium; cell fusion; cell membrane; chloride channels; egg /ovum; electrophysiology; fertilization; fluorescence microscopy; immunocytochemistry; in situ hybridization; meiosis; membrane proteins; microinjections; receptor; sea urchins; serotonin; sperm; western blottings

To investigate the signals that pass from the sperm to the egg to initiate development, the function of G-proteins and G-protein-linked receptors in the egg membrane will be examined. The specific aims are: 1) To identify and characterize G-protein alpha-subunits in eggs and embryos. 2) To investigate how closely G-protein stimulation mimics events at fertilization. 3) To investigate the effects of inhibiting G-protein function in eggs, as well as in oocytes and embryos. 4) To determine if G- proteins are activated at fertilization. 5) To investigate the functions of G-protein-linked receptors in eggs, as well as in oocytes and embryos. The experimental design and methods include immunoblotting of egg membranes with G-protein-specific antibodies, measurements of calcium transients and calcium-mediated ion channel opening, microinjection of antibodies against particular G-proteins, use of azido-anilido-GTP to detect G-protein activation, and expression in eggs of mRNAs encoding G-protein-linked receptors. The significance of this work to the study of human development is that the cell biological principles we discover in our work with echinoderms and amphibians will form a basis for future studies of mammals. This basic science background may ultimately find applications to practical problems such as contraception and infertility.

研究从精子传递到卵子的信号以启动 发育、G 蛋白和 G 蛋白连接受体的功能 将检查卵膜。 具体目标是： 1) 确定 并表征卵子和胚胎中的 G 蛋白 α 亚基。 2) 至 研究 G 蛋白刺激与事件的模拟程度如何 施肥。 3) 研究抑制G蛋白的作用 在卵子、卵母细胞和胚胎中发挥作用。 4) 判断是否G- 蛋白质在受精时被激活。 5）研究功能 卵子、卵母细胞和胚胎中的 G 蛋白连接受体。 实验设计和方法包括卵膜免疫印迹 使用 G 蛋白特异性抗体，测量钙瞬变和 钙介导的离子通道开放，显微注射抗体 特定的 G 蛋白，使用叠氮基苯胺基 GTP 检测 G 蛋白 编码 G 蛋白连接的 mRNA 在卵中的激活和表达 受体。 这项工作对人类发展研究的意义 是我们在工作中发现的细胞生物学原理 棘皮动物和两栖动物将成为未来哺乳动物研究的基础。 这种基础科学背景最终可能会应用于实际 避孕和不孕等问题。