GENE-TARGETED MODULATION OF BRAIN DOPAMINE SYSTEMS

描述：(改编自申请者摘要)总体目标 这个研究方案的目的是使用有针对性的表达方式 控制大脑多巴胺能系统的异源基因。这 一项提案利用了最近对cdna的克隆 神经递质转运体。两个考虑因素包括 这项工作的基本原理。首先，现有数据表明， 多巴胺转运体(DAT)仅在多巴胺能神经元中表达。 事实上，多巴胺转运蛋白可能是唯一一种其 表达仅限于多巴胺能神经元。因此，发起人 多巴胺转运体的区域将是分析的独特工具 多巴胺系统的功能，以及一个潜在的基因工具 治疗，因为它将可能靶向表达一种 异源蛋白选择性地与多巴胺能神经元结合。第二 考虑的是多巴胺系统在各种疾病中的重要性 精神分裂症、帕金森症和药物滥用等疾病。这个 该提案的具体目标基于两个指导性假设：1) 多巴胺转运体的细胞特异性表达由以下因素决定 基因5‘侧翼区的顺式作用元件。这个 拟议的实验还将确定关键的监管要素 都不在5‘侧翼区域。2)它将是 有可能定义多巴胺的转录调控结构域 针对异源基因强表达的转运蛋白基因 蛋白质选择性地作用于多巴胺能神经元。该计划的具体目标是 应用是确定决定多巴胺能的基因组元件 转基因小鼠的细胞特异性表达。

DESCRIPTION: (adapted from Applicant's Abstract) The overall objective of this research program is to use the targeted expression of heterologous genes to manipulate brain dopaminergic systems. This proposal takes advantage of the recent cloning of the cDNA for several neurotransmitter transporters. Two considerations comprise the rationale for this work. First, the available data indicate that the dopamine transporter (DAT) is expressed only in dopaminergic neurons. Indeed, the dopamine transporter may be the only protein whose expression is limited to dopaminergic neurons. Thus, the promoter region of the dopamine transporter will be a unique tool for the analysis of the function of the dopamine system, and a potential tool for gene therapy, because it will be possible to target expression of a heterologous protein selectively to dopaminergic neurons. The second consideration is the importance of the dopamine system in a variety of disorders including schizophrenia, parkinsonism and drug abuse. The specific aims of this proposal are based on two guiding hypotheses: 1) Cell specific expression of the dopamine transporter is determined by cis-acting elements in the 5' flanking region of the gene. The experiments proposed will also determine if critical regulatory elements are present that are not in the 5' flanking region. 2) It will be possible to define a transcriptional regulatory domain of the dopamine transporter gene that will target strong expression of a heterologous protein selectively to dopaminergic neurons. The specific aim of the application is to identify genomic elements that determine dopaminergic cell-specific expression in transgenic mice.