Molecular and Genetic Analysis of Presynaptic Proteins

描述(申请人提供)：我们的目标是继续我们对土壤线虫秀丽线虫突触传递的分子和遗传分析。线虫之所以被用于这些研究，是因为它的神经系统简单，易于进行遗传和分子分析，以及其基因组的完整测序。这项拟议的研究是基于最近的结果和最新的技术进步。这三个主要目标是：(1)深入分析最近描述的rip-1基因，它编码一种极大的蛋白质，被认为是为突触前神经末梢的组成部分提供“分子支架”；(2)开始研究神经终端内用来调节包装到每个突触小泡中的神经递质的数量的机制；以及(3)继续使用线虫细胞培养系统来分析突触的体外发育和功能。拟议的研究将扩展我们之前对土壤线虫线虫突触结构和神经递质功能的分析，并将包括分离缺乏囊泡含量调节的突变体，以及鉴定适当调节量子大小所需的特定分子和信号通路。虽然这是基础研究，但它显然与健康有关，因为适当的神经递质释放对神经系统的正常功能至关重要，而且递质释放缺陷与许多精神和神经疾病有关。

DESCRIPTION (provided by applicant): It is our goal to continue our molecular and genetic analysis of synaptic transmission in the soil nematode Caenorhabditis elegans. C. elegans is being used for these studies because of its simple nervous system, its ease of genetic and molecular analysis, and the complete sequencing of its genome. The proposed research is based on recent results and on recent technological advances. The three broad goals are: (1) to analyze in depth the recently-described rip-1 gene, which encodes an extremely large protein thought to provide a "molecular scaffold" for the components of a presynaptic nerve terminal; (2) to begin an investigation of the mechanisms used within nerve terminals to regulate the amount of neurotransmitter packaged into each synaptic vesicle; and (3) to continue studies using a C. elegans cell culture system for the analysis of synaptic development and function in vitro. The proposed studies will extend our previous analysis of synaptic structure and neurotransmitter function in the soil nematode Caenorhabditis elegans, and will include the isolation of mutants deficient in the regulation of vesicle content, and the identification of specific molecules and signaling pathways required for proper modulation of quantal size. Although this is basic research, it is clearly relevant to health, since proper neurotransmitter release is crucial to proper function of the nervous system, and defective transmitter release has been implicated in many psychiatric and neurological disorders.