CELL BIOLOGY OF THE SYNAPSE

突触的细胞生物学

• 批准号: 3394959
• 负责人: PHILIP SIEKEVITZ
• 金额: $ 23.27万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-05-01 至 1986-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3394959
• 关键词: affinity chromatography; autoradiography; calcium; cell membrane; cyclic AMP; density gradient ultracentrifugation; electron microscopy; electrophysiology; gel electrophoresis; immunochemistry; immunodiffusion; membrane structure; muscle proteins; neural conduction; neural transmission; neurochemistry; neuromuscular junction; neurotransmitters; phosphorylation; radiotracer; scanning electron microscopy; smooth muscle; spider poison; synapses; synaptic vesicles; synaptosomes; tissue /cell culture

The function of calmodulin and of the major 51,000 Mr protein in cerebral cortex postsynaptic densities (PSD), and the physiological significance of these funtions, will be examined in two ways. First, antibodies to these two proteins will be injected into neuronal cells by micromanipulation, and the effects of these will be monitored electrophysiologically, under normal conditions and under depolarization conditions. Second, a spin-label will be attached to calmodulin, and the spin-labeled calmodulin reconstituted into PSDs of a synaptosome preparation depleted of calmodulin. The synaptosome preparation will be depolarized in various ways, and the ESR signals recorded to observe any change in the movement of the calmodulin during the post-synaptic response. In another project, an attempt will be made to purify alpha-latrotoxin from American black widow spiders, since the biochemical procedures used for the Italian spiders were unsuccessful. This attempt will be based on the strong specific binding of the toxin to synaptic membranes and the removal of the toxin from the membranes by antibodies to it.

大脑皮层钙调素和主要51，000 Mr蛋白的功能 突触后密度（PSD），以及这些的生理意义 功能，将从两个方面进行研究。 首先，针对这两种蛋白质的抗体 将通过显微操作注射到神经细胞中， 这些将在正常条件下进行电生理监测， 在去极化条件下。 第二，一个自旋标签将附加到 钙调蛋白，自旋标记的钙调蛋白重组成PSD的 突触体制备耗尽钙调蛋白。 突触体制备 将以各种方式去极化，并记录ESR信号以观察任何 突触后反应期间钙调素运动的变化。 在 另一个项目，将尝试从 美国黑寡妇蜘蛛，因为生化程序用于 意大利蜘蛛没有成功。 这一尝试将以强者为基础 毒素与突触膜的特异性结合以及毒素的去除 从细胞膜中分离出来