MOLECULAR AND GENETIC ANALYSIS OF PRESYNAPTIC PROTEINS

我们的目标是继续我们的分子和遗传分析突触 在土壤线虫秀丽隐杆线虫中传播。 我们有 确定了一组似乎编码突触前蛋白的基因; 当这些基因突变时，会导致过量乙酰胆碱的积累， 还赋予对乙酰胆碱酯酶抑制剂的抗性。 一些 这些基因被证明编码重要的突触囊泡蛋白 如突触结合蛋白和囊泡乙酰胆碱转运蛋白。 另一个基因，称为unc-41最近被克隆，似乎 编码一种新的突触前蛋白。 这件事背后的主题 研究是识别新的突触前蛋白， 确定其功能。拟议的三大目标 研究内容包括：（1）鉴定影响突触前功能的新基因 通过对一系列突变体的遗传和表型分析， 乙酰胆碱酯酶抑制剂耐药;（2）继续分子 unc-41基因和nc-41蛋白的分析;和（3）开始 对新发现的突触前基因进行了详细的分子研究。 C.秀丽隐杆线虫被用于这些研究，因为它简单的神经 系统，其易于遗传和分子分析，以及可用性 利用DNA介导的转化技术导入克隆基因 这个有机体。 对unc-41的研究旨在了解 突触传递中的作用。的信息 获得的关于野生型基因结构和转录本将用于 分析一系列UNC-41突变体 针对 将生产UNC-41蛋白并用于免疫定位研究。 突变将在与unc-41相互作用的基因中被分离出来， 突变可能编码额外的突触前成分。 的 对unc-41的调查将为分析 额外的突触前基因 虽然这是基础研究， 显然与健康有关，因为适当的神经递质释放是 对神经系统的正常功能至关重要， 递质释放与许多精神疾病有关， 神经系统疾病

It is our goal to continue our molecular and genetic analysis of synaptic transmission in the soil nematode Caenorhabditis elegans. We have identified a set of genes which appear to encode presynaptic proteins; these genes, when mutated, cause accumulation of excess acetylcholine and also confer resistance to inhibitors of acetylcholinesterase. Some of these genes have been shown to encode important synaptic vesicle proteins such as synaptotagmin and the vesicular acetylcholine transporter. Another gene, called unc-41 has recently been cloned and appears to encode a novel presynaptic protein. The underlying theme of this research is the identification of new presynaptic proteins and the determination of their function. The three broad goals of the proposed research are: (l) to identify new genes affecting presynaptic function through genetic and phenotypic analysis of a collection of mutants resistant to acetylcholinesterase inhibitors; (2) continue the molecular analysis of the unc-41 gene and the UNC-41 protein; and (3) begin detailed molecular studies on the newly identified presynaptic genes. C. elegans is being used for these studies because of its simple nervous system, its ease of genetic and molecular analysis, and the availability of the technique of DNA-mediated transformation to introduce cloned genes into this organism. The studies on unc-41 are aimed at understanding the role of the UNC-41 protein in synaptic transmission. The information obtained about wild-type gene structure and transcripts will be used to analyze a collection of unc-41 mutants. Antibodies directed against the UNC-41 protein will be produced and used for immunolocalization studies. Mutations will be isolated in genes which interact with unc-41; such mutations might encode additional presynaptic components. The investigations of unc-41 will provide a paradigm for the analysis of additional presynaptic genes. 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.