INTEGRINS AND EGG ACTIVATION AT FERTILIZATION

这项研究的总体目标是确定整合素 在受精时参与卵子激活的过程。这将是 以多种方式接受调查。第一，存在和本土化 在α整合素亚基中，可能参与的整合素亚基 在两栖类受精过程中，将使用免疫印迹和 免疫荧光技术。将寻找特定的Beta亚基 使用免疫沉淀和免疫印迹。第二，职能作用 对于受精时的整合素，将使用功能阻断进行检测 抗整合素亚基抗体干扰精-卵融合 和/或卵子激活。第三个目标是确定是否 激活鸡蛋表面的整合素刺激鸡蛋的活动 活化，包括产生磷脂酰肌醇4，5- 双磷酸盐，膜电位变化，细胞内增加 钙离子，受精包膜的升高，皮质收缩， 第二极体的形成和肌动蛋白在 精子进入。最终目标是为 推测哺乳动物整合素精子受体进入非洲爪哇卵子并 检查它们的表达是否允许小鼠精子与青蛙融合 卵子并启动卵子激活。这些研究将有助于我们的 了解受精的基本生物学知识。这一背景可能 最终发现在避孕等实际问题上的应用 和不孕不育。

The overall objective of this research is to determine if integrins participate in the process of egg activation at fertilization. This will be investigated in a number of ways. First, the presence and localization of the alpha integrin subunit, an integrin subunit likely to participate in amphibian fertilization, will be determined using immunoblotting and immunofluorescence techniques. A specific Beta subunit will be looked for using immunoprecipitation and immunoblotting. Second, a functional role for integrins at fertilization will be examined using function-blocking antibodies against integrin subunits to interfere with sperm-egg fusion and/or egg activation. The third objective is to determine whether activating integrins on the egg surface stimulates events of egg activation, including production of phosphatidylinositol 4,5- bisphosphate, changes in membrane potential, an increase in intracellular Ca2+, elevation of the fertilization envelope, cortical contraction, second polar body formation, and actin accumulation in the region of sperm entry. The final objective is to introduce the subunits for the putative mammalian integrin sperm receptor into Xenopus eggs and to examine whether their expression allows mouse sperm to fuse with frog eggs and initiate egg activation. These studies will contribute to our understanding of the basic biology of fertilization. This background may ultimately find applications to practical problems such as contraception and infertility.