MOLECULAR BIOLOGY OF THE SYNAPSE

突触的分子生物学

基本信息

批准号：
6637660
负责人：
EILEEN M. LAFER
金额：
$ 32.51万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
1987
资助国家：
美国
起止时间：
1987-04-01 至 2004-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6637660
关键词：
Caenorhabditis elegans binding sites clathrin endocytosis molecular biology neural transmission phosphoproteins protein binding protein protein interaction protein structure function receptor binding squid synapses synaptic vesicles synaptotagmin tissue /cell culture

项目摘要

DESCRIPTION (Adapted from applicant's abstract): The long term goal of this project is to determine the molecular mechanisms that underlie neurotransmission. The goal of the next project period is to understand the role of clathrin-mediated vesicle recycling in neurotransmission as well as to better understand the molecular mechanisms of clathrin-coat assembly and disassembly. Aim 1 tests the hypothesis that synaptic vesicle coating and uncoating are required for normal neurotransmission by examining the effects of reagents that disrupt clathrin assembly and vesicle uncoating on neurotransmission in the squid giant synapse preparation. Aim 2 will determine whether synaptic vesicle proteins participate in the formation of clathrin coats. A combination of biochemical and physiological techniques will be used to critically evaluate the proposed roles of the synaptic vesicle proteins synaptotagmin, synaptobrevin and SV2 as well as a novel protein identified in a yeast two hybrid screen for proteins that bind the clathrin adaptor protein AP180. Aim 3 will assess the physiological importance of two regions of the adaptor protein AP180, the clathrin assembly domain and the inositol lipid-binding domain, by testing these activities in C. elegans AP180 mutant proteins and by assessing the ability of novel mutant proteins to rescue and AP180 mutant phenotype. Aim 4 will critically evaluate the hypothesis that interactions between resident coat proteins regulate the efficiency of synaptic vesicle endocytosis utilizing a combination of biochemical and genetic approaches. Studies will focus on interactions between AP180, AP2, eps15 and auxilin. Aim 5 will biochemically characterize novel C. elegans AP180 mutants that have defects in synaptic transmission. Successful completion of these studies will allow us to achieve a deeper level of understanding of the processes of endocytosis and neurotransmission.

描述（改编自申请人的摘要）：本项目的长期目标项目的目的是确定其背后的分子机制神经传递。下一个项目期的目标是了解网格蛋白介导的囊泡回收在神经传递中的作用以及更好地了解网格蛋白涂层组装的分子机制拆卸。目标 1 检验突触小泡涂层和通过检查以下因素的影响，正常的神经传递需要脱涂层破坏网格蛋白组装和囊泡脱壳的试剂鱿鱼巨突触制备中的神经传递。目标 2 将决定突触小泡蛋白是否参与网格蛋白的形成外套。将结合使用生化和生理技术严格评估突触小泡蛋白的拟议作用 synaptotagmin、synaptobrevin 和 SV2 以及在酵母两种杂交筛选结合网格蛋白衔接蛋白的蛋白质 AP180。目标 3 将评估大脑两个区域的生理重要性接头蛋白 AP180、网格蛋白组装结构域和肌醇脂质结合域，通过测试秀丽隐杆线虫 AP180 突变体中的这些活性蛋白质并通过评估新型突变蛋白拯救和 AP180突变表型。目标 4 将批判性地评估以下假设：常驻外壳蛋白之间的相互作用调节突触的效率利用生化和遗传相结合的囊泡内吞作用接近。研究将重点关注 AP180、AP2、eps15 和辅助素。目标 5 将对新型线虫 AP180 突变体进行生化表征突触传递有缺陷。顺利完成这些研究将使我们对事物有更深入的了解内吞作用和神经传递过程。