MOUSE SYNAPTIC VESICULAR MONAMINE TRANSPORTER--CDNA AND GENOMIC STRUCTURE

小鼠突触小泡单胺转运蛋白--CDNA和基因组结构

• 批准号: 2571614
• 负责人: G R UHL
• 金额: --
• 依托单位: NATIONAL INSTITUTE ON DRUG ABUSE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2571614
• 关键词: amphetamines; animal genetic material tag; complementary DNA; dopamine; dopamine transporter; embryonic stem cell; gene deletion mutation; genetic regulatory element; laboratory mouse; methylphenyltetrahydropyridine; molecular cloning; neurotoxicology; neurotoxins; neurotransmitter transport; nucleic acid sequence; synaptic vesicles; transcription factor; transfection /expression vector

Activities of the principal brain vesicular monamine transporter, VMAT2, are key to understanding the cellular compartmentalization of monoamines that may play a key role in modulating the actions and neurotoxicities induced by amphetamine and related psychostimulants. In previous FYs, investigators in this Branch identified cDNAs encoding human VMAT2, and cloned murine VMAT2 cDNA and genomic DNA clones from ES cell lines and to define the VMAT2 genomic structure and major transcriptional start sites for the VMAT2 gene. Sequence analysis of 5' flanking sequences revealed a putative promoter region containing several consensus sequences for transcription factor binding. Analyses of these cDNA and genomic clones has allowed construction of targeting vectors for deletion of several VMAT2 exons in homologous recombinant Aknockout mice. Homozygous knockouts die shortly after birth, but heterozygous mice display differences in responses to amphetamine reward and locomotion that point strongly toward differential impact of amphetamine-induced vesicular release on these two processes. They also display altered sensitivity to MPP+ dopaminergic toxicity. These mice substantially enhance our understanding of mechanisms of psychostimulant and dopaminergic neurotoxin action.

主要脑囊泡单胺转运蛋白VMAT 2的活性， 是理解单胺细胞区室化的关键 可能在调节神经毒性的作用中起着关键作用 安非他明和相关精神兴奋剂引起的。 在以前的财政年度， 该分支的研究者鉴定了编码人VMAT 2的cDNA， 从ES细胞系克隆鼠VMAT 2 cDNA和基因组DNA克隆， 为了确定VMAT2基因组结构和主要转录起始点， VMAT2基因的位点。 5'侧翼序列的序列分析 揭示了一个假定的启动子区域， 用于转录因子结合的序列。 这些cDNA的分析和 基因组克隆允许构建用于缺失的靶向载体 同源重组A基因敲除小鼠中几个VMAT2外显子的表达。 纯合子敲除小鼠出生后不久死亡，但杂合子小鼠 对安非他明奖励和运动的反应 这有力地表明，安非他明引起 在这两个过程中的囊泡释放。 它们还显示出 对MPP+多巴胺能毒性敏感。 这些老鼠基本上 提高我们对精神兴奋剂机制的理解， 多巴胺能神经毒素作用。