MOLECULAR AND GENETIC ANALYSIS OF PRESYNAPTIC PROTEINS

我们的目标是继续我们对突触的分子和基因分析 线虫在土壤中的传播。我们有 确定了一组似乎编码突触前蛋白的基因； 当这些基因发生突变时，会导致过量的乙酰胆碱和 对乙酰胆碱酯酶抑制剂也有抵抗力。其中一些 这些基因已被证明编码重要的突触小泡蛋白。 如突触素和囊泡乙酰胆碱转运蛋白。 另一种名为UNC-41的基因最近被克隆，似乎 编码一种新的突触前蛋白。这件事的基本主题 研究正在鉴定新的突触前蛋白和 它们的功能的确定。建议的三个主要目标 研究内容有：(L)寻找影响突触前功能的新基因 通过对一组突变体的遗传和表型分析 抗乙酰胆碱酯酶抑制剂；(2)继续分子 UNC-41基因和UNC-41蛋白的分析；和(3)开始 对新发现的突触前基因的详细分子研究。 线虫之所以被用于这些研究，是因为它简单而紧张 系统，它的遗传和分子分析的简便性，以及可用性 DNA介导法导入克隆基因的研究进展 进入这个有机体。对UNC-41的研究旨在了解 UNC-41蛋白在突触传递中的作用。这些信息 获得的有关野生型基因结构和转录本将用于 分析一组UNC-41突变体。针对该病毒的抗体 UNC-41蛋白将被生产并用于免疫定位研究。 突变将被隔离在与UNC-41相互作用的基因中； 突变可能编码额外的突触前成分。这个 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，但它是 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，且有缺陷 递质释放已经牵涉到许多精神病和 神经紊乱。

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.