MOLELULAR AND GENETIC ANALYSIS OF PRESYNAPTIC PROTEINS

突触前蛋白的分子和遗传分析

• 批准号: 2271820
• 负责人: James B. Rand
• 金额: $ 18.04万
• 依托单位: OKLAHOMA MEDICAL RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2271820
• 关键词: Caenorhabditis elegans; acetylcholine; acetylcholinesterase; alleles; artificial chromosomes; chickens; cholinesterase inhibitors; complementary DNA; drug resistance; immunocytochemistry; laboratory rabbit; nerve /myelin protein; neuropharmacology; neurotransmitter transport; pharmacogenetics; sequence tagged sites; suppressor mutations; synaptic vesicles; transfection /expression vector; yeasts

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.

我们的目标是继续我们的突触分子和遗传分析 土壤中的传播线虫秀丽隐杆线虫。 我们有 确定了一组似乎编码突触前蛋白的基因； 这些基因在突变时会导致过量的乙酰胆碱和 还赋予对乙酰胆碱酯酶抑制剂的耐药性。 其中一些 这些基因已被证明编码重要的突触囊泡蛋白 例如突触毒素和囊泡乙酰胆碱转运蛋白。 另一个称为unc-41的基因最近被克隆了，似乎 编码一种新型突触前蛋白质。 这个基本主题 研究是鉴定新的突触前蛋白质和 确定其功能。拟议的三个广泛目标 研究是：（l）确定影响突触前功能的新基因 通过对突变体集合的遗传和表型分析 对乙酰胆碱酯酶抑制剂的抗性； （2）继续分子 分析UNC-41基因和UNC-41蛋白； （3）开始 关于新鉴定的突触前基因的详细分子研究。 秀丽隐杆线虫因其简单的神经而被用于这些研究 系统，遗传和分子分析的易用性以及可用性 DNA介导的转化技术引入克隆基因 进入这个生物体。 关于UNC-41的研究旨在了解 UNC-41蛋白在突触传播中的作用。信息 关于野生型基因结构和转录本获得的 分析一组UNC-41突变体。 针对的抗体 UNC-41蛋白将用于免疫定位研究。 突变将在与UNC-41相互作用的基因中分离出来。这样的 突变可能编码其他突触前成分。 这 对UNC-41的调查将为分析提供范式 其他突触前基因。 尽管这是基础研究，但这是 显然与健康有关，因为适当的神经递质释放是 对于神经系统的适当功能至关重要，有缺陷 发射器释放已与许多精神科有关， 神经系统疾病。