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: 6161705
• 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/6161705
• 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. In this FY, they completed work cloning murine VMAT2 cDNA and genomic DNA clones from ES cell lines and defining VMAT2 genomic structure and transcriptional initiation sites. 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 "knockout" 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.

主要脑泡单胺转运蛋白的活动， VMAT2 是理解细胞区室化的关键 单胺可能在调节作用和 安非他明和相关精神兴奋剂引起的神经毒性。 在之前的财政年度中，该部门的研究人员鉴定了编码 人类 VMAT2。在本财年，他们完成了小鼠 VMAT2 cDNA 的克隆工作 和来自 ES 细胞系的基因组 DNA 克隆并定义 VMAT2 基因组 结构和转录起始位点。 序列分析 5' 侧翼序列揭示了一个推定的启动子区域，其中包含 转录因子结合的几个共有序列。 分析 这些 cDNA 和基因组克隆允许构建靶向 用于在同源重组中删除多个 VMAT2 外显子的载体 “淘汰”小鼠。 纯合子基因敲除在出生后不久就会死亡，但是 杂合子小鼠对安非他明的反应表现出差异 奖励和运动强烈表明不同的影响 安非他明诱导的囊泡释放对这两个过程的影响。 他们 还表现出对 MPP+ 多巴胺能毒性的敏感性改变。 这些 小鼠大大增强了我们对机制的理解 精神兴奋和多巴胺能神经毒素作用。