SIGNAL TRANSDUCTION AT FERTILIZATION

受精时的信号转导

• 批准号: 2197144
• 负责人: LAURINDA A. JAFFE
• 金额: $ 11万
• 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/2197144
• 关键词: Asteroidea; DNA replication; G protein; Rana; Xenopus oocyte; alternatives to animals in research; biological signal transduction; calcium; cell fusion; cell membrane; chloride channels; egg /ovum; electrophysiology; fertilization; fluorescence microscopy; immunocytochemistry; in situ hybridization; meiosis; membrane proteins; microinjections; nucleic acid sequence; 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.

调查从精子到鸡蛋传递的信号以启动 开发，G蛋白和G蛋白连接受体的功能 将检查鸡蛋膜。 具体目的是：1）确定 并表征卵和胚胎中的G蛋白α-亚基。 2）到 研究如何在G蛋白刺激模仿事件中如何接近 受精。 3）研究抑制G蛋白的影响 在鸡蛋以及卵母细胞和胚胎中的功能。 4）确定g-是否 蛋白质在受精时被激活。 5）调查功能 卵中的G蛋白连接受体，卵母细胞和胚胎中的受体。 实验设计和方法包括卵膜的免疫印迹 用G蛋白特异性抗体，钙瞬变的测量和 钙介导的离子通道开口，对抗体的显微注射 特定的G蛋白，使用叠氮化杂氮-GTP检测G蛋白 激活和在编码G蛋白连接的mRNA卵中的表达 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为将来对哺乳动物进行研究的基础。 这种基础科学背景最终可能会找到实用的申请 避孕和不育等问题。