ANESTHETIC MECHANISMS IN GABAA R GENE TARGETED MICE

GABAA R 基因靶向小鼠的麻醉机制

• 批准号: 6019045
• 负责人: LEONARD L FIRESTONE
• 金额: $ 39.39万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-01-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6019045
• 关键词: GABA receptor; anesthetics; biotechnology; electrophysiology; gene targeting; general anesthesia; genetically modified animals; histology; laboratory mouse; method development; neuropharmacology; point mutation; tissue /cell culture; transfection

DESCRIPTION (Adapted from applicant's abstract): This proposal is focused on determination of the molecular mechanisms of general anesthetics using transgenic techniques. The investigators will focus on the type A-aminobutryic acid receptor. Mouse lines that ubiquitously lack either the beta3, 26, or delta subunits of the GABAA-R have been produced and studies are underway in these cells/animals. In addition, the researchers proposed to create four further novel mouse lines that harbor tissue-specific knockouts of the GABAA-R2 or beta3 subunits. This conditional knockout approach has major advantages in that untoward developmental effects of the genetic alteration are avoided and the technique permits analysis of the putative neuroanatomic site of action of the anesthetic agents. Finally, the investigators will examine the anesthetic response of three additional genetically altered mouse lines: a) ubiquitous knockout of the alpha1, subunit of the GABAA-R; b) a transgene that expresses an Ala291 Trp point mutation in the alpha1 subunit; c) a knockin mutation of the Ala291 Trp mutation that replaces the endogenous alpha1 gene.

描述（改编自申请人摘要）：本提案的重点 关于确定全身麻醉药的分子机制， 转基因技术 调查人员将重点调查 A-氨基丁酸受体 鼠标线，普遍缺乏 GABAA-R的β 3、26或δ亚基已被生产和研究 正在这些细胞/动物中进行。 此外，研究人员还提出， 创造出四种新的小鼠品系， GABAA-R2或β 3亚基的敲除。 这种条件性击倒 这种方法的主要优点在于， 避免了遗传改变，并且该技术允许分析 麻醉剂作用的假定神经解剖学部位。 最后， 研究人员将检查另外三个人的麻醉反应， 遗传改变的小鼠系：a）α 1的普遍敲除， GABAA-R的亚基; B）表达Ala 291 Trp点的转基因 α 1亚基中的突变; c）Ala 291 Trp的敲入突变 替代内源性α 1基因的突变。