SIGNAL TRANSDUCTION AT FERTILIZATION

研究从精子传递到卵子的信号以启动 G蛋白和G蛋白相关受体的发育、功能 将检查卵膜。具体目标是：1)识别 并对鸡蛋和胚胎中的G蛋白α亚基进行了表征。2)至 研究G蛋白刺激在多大程度上模拟事件 受精。3)研究抑制G蛋白的作用 在卵子以及卵母细胞和胚胎中的功能。4)确定G- 蛋白质在受精时被激活。5)研究功能 在卵子中，以及在卵母细胞和胚胎中，G蛋白连接的受体的数量。 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，微量注射抗 用叠氮-苯胺-GTP检测G-蛋白 编码G蛋白的mRNAs的激活及其在鸡蛋中的表达 感受器。这项工作对研究人类发展的意义 我们在研究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。 这一基础科学背景最终可能会在实际应用中 避孕和不孕不育等问题。

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.