GENETIC REGULATION OF ACETYLCHOLINE SYNTHESIS

我们的目标是继续我们对CHA-1的分子和基因分析 土壤线虫秀丽线虫基因复合体。CHA-1是 胆碱乙酰转移酶(ChAT)的结构基因 合成神经递质乙酰胆碱。以前获得的 有证据表明，线虫体内的Chat蛋白可能包括 多个结构和功能域，我们对 确定基因的组织。此外，聊天被认为是一种 胆碱能神经元分化特异性标记物，我们希望 研究其在神经发育过程中的空间和时间控制。C。 秀丽被用于这些研究，因为它简单而紧张 系统，其遗传分析的简便性，以及该技术的可用性 从这种有机体中克隆基因。我们最近做了 使用转座元件TCL来标记和克隆CHA-1的一部分 很复杂。将构建基因的物理图谱，并将其关联起来 基因图谱已经可用。该病毒的完整核苷酸序列 然后，基因将被确定。该地区的成绩单将是 进行了Northern分析、S1分析和cDNA的分离 序列。这样构建的转录图谱将与 基因图谱；这将测试我们发布的聊天结构模型和 功能。获得的关于野生型基因结构和 然后，转录本将被用于分析胆碱能缺陷突变 功能。将产生针对Chat的抗体，用于 生化和解剖学研究。虽然这是基础研究，但它是 显然与健康有关，因为乙酰胆碱代谢的变化 和/或胆碱能功能已在许多神经病学和 精神障碍，包括阿尔茨海默病、肌萎缩侧索硬化症和抑郁症。

It is our goal to continue our molecular and genetic analysis of the cha-1 gene complex from the soil nematode Caenorhabditis elegans. cha-1 is the structural gene for choline acetyltransferase (ChAT), the enzyme which synthesizes the neurotransmitter acetylcholine. Previously obtained evidence suggests that the ChAT protein in C. elegans may consist of multiple structural and functional domains, and we are interested in determining the organization of the gene. In addition, ChAT is considered a differentiation-specific marker for cholinergic neurons, and we wish to investigate its spatial and temporal control during neural development. C. elegans is being used for these studies because of its simple nervous system, its ease of genetic analysis, and the availability of the technique of "transposon tagging" to clone genes from this organism. We have recently used the transposable element Tcl to "tag" and clone part of the cha-1 complex. A physical map of the gene will be constructed, and correlated with the genetic map already available. The complete nucleotide sequence of the gene will then be determined. Transcripts from the region will be studied by Northern analysis, S1 analysis, and the isolation of cDNA sequences. The transcriptional map thus constructed will be compared to the genetic map; this will test our published model of ChAT structure and function. The information obtained about wild-type gene structure and transcripts will then be used to analyze mutants defective in cholinergic function. Antibodies directed against ChAT will be produced, for use in biochemical and anatomical studies. Although this is basic research, it is clearly relevant to health, since alterations in acetylcholine metabolism and/or cholinergic function have been identified in many neurological and psychiatric disorders, including Alzheimer's disease, ALS, and depression.