GENE-TARGETED MODULATION OF BRAIN DOPAMINE SYSTEMS

描述：（改编自申请人的摘要）总体目标 这项研究计划的目的是利用靶向表达 异源基因来操纵大脑多巴胺能系统。 这 该提案利用了最近克隆的几个cDNA， 神经递质转运蛋白 两个考虑因素包括 这项工作的理由。 首先，现有数据表明， 多巴胺转运蛋白（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 表达限于多巴胺能神经元。 因此，启动子 多巴胺转运蛋白的区域将是分析的独特工具， 多巴胺系统的功能，也是基因治疗的潜在工具。 治疗，因为它将有可能靶向表达的一个 异源蛋白选择性地作用于多巴胺能神经元。 第二 考虑的是多巴胺系统在各种 包括精神分裂症、帕金森综合症和药物滥用在内的疾病。 的 本建议的具体目标基于两个指导假设： 多巴胺转运蛋白的细胞特异性表达由以下决定： 在基因的5'侧翼区域中的顺式作用元件。 的 提出的实验也将确定是否关键的调控元件 不存在于5'侧翼区中。 2)将 有可能定义多巴胺的转录调控域 转运蛋白基因，其将靶向强表达异源 蛋白质选择性地作用于多巴胺能神经元。 的具体目标 应用是确定决定多巴胺能的基因组元件， 转基因小鼠中的细胞特异性表达。

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.