SIGNAL TRANSDUCTION AT FERTILIZATION

描述（改编自研究者摘要）：本次更新 该应用涉及精子的信号机制 使卵子在受精时开始发育。 该项目 集中在这个过程的最早期， 质膜 这些膜事件导致细胞内Ca 2+升高， 这反过来又导致了许多后续的发展过程。 项目 包括3个具体目标：（1）研究G-α-q家族在肿瘤中的作用 蛋白质在受精信号转导中的作用 G-α-q的含量 通过注射反义核酸， 寡核苷酸，以确定这是否抑制Ca 2+释放， 受精 这些研究还将检查G-α-q是否是 在受精时激活，利用G-α-q蛋白的保护作用， 蛋白水解作为一种分析。 (2)研究酪氨酸的作用 信号转导中磷脂酶C-γ的磷酸化. 受精 磷脂酶C-γ的激活将被抑制， 注射PLC-γ SH 2结构域，以确定这是否减少Ca 2 + 受精时释放。 这些研究还将检查PLC-γ是否 在受精时被激活，利用这种酶的酪氨酸磷酸化 作为分析。 (3)为了研究α-v整合素在信号转导中的作用， 受精时的转导。 这些研究将调查是否 α-v整联蛋白的刺激引起细胞内Ca 2+升高， 两栖类卵，以及它是否增加脂质PIP 2的量， 磷脂酶C底物。他们还将检查Ca 2+释放是否 通过抑制α-V整联蛋白功能来降低AT受精。

DESCRIPTION (Adapted from the Investigator's Abstract): This renewal application is concerned with the signalling mechanisms by which a sperm causes an egg to begin development at fertilization. The project is focussed on the earliest events of this process, that occur at the egg plasma membrane. These membrane events cause a rise in intracellular Ca2+, which in turn causes many subsequent developmental process. The project includes 3 specific aims: (1) To examine the role of G-alpha-q family proteins in signal transduction at fertilization. The amounts of G-alpha-q family proteins in frog eggs will be reduced by injection of antisense oligonucleotides, to determine if this inhibits Ca2+ release at fertilization. These studies will also examine whether G-alpha-q is activated at fertilization, using protection of the G-alpha-q protein from proteolysis as an assay. (2) To examine the role of tyrosine phosphorylation of phospholipase C-gamma in signal transduction at fertilization. Activation of phospholipase C-gamma will be inhibited by injection of PLC-gamma SH2 domains, to determine if this reduces Ca2+ release at fertilization. These studies will also examine whether PLC-gamma is activated at fertilization, using tyrosine phosphorylation of this enzyme as a assay. (3) To examine the role of alpha-v integrins in signal transduction at fertilization. These studies will investigate whether stimulation of an alpha-v integrin causes a rise in intracellular Ca2+ in amphibian eggs, and whether it increases the amount of the lipid PIP2, the substrate for phospholipase C. They will also examine whether Ca2+ release at fertilization is reduced by inhibition of alpha-v integrin function.