BRAIN MEMBRANES IN ALCOHOL AND BENZODIAZEPINE DEPENDENCE

酒精和苯二氮卓依赖的脑膜

• 批准号: 2043455
• 负责人: Robert A Harris
• 金额: $ 24.98万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-09-29 至 1999-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2043455
• 关键词: Xenopus oocyte; alcoholism /alcohol abuse; barbiturates; benzodiazepines; brain cell; brain metabolism; calcium; chloride channels; drug addiction; drug interactions; drug metabolism; drug tolerance; electrophysiology; fluorescence spectrometry; fluorescent dye /probe; gene expression; genetically modified animals; inositol phosphates; laboratory mouse; liver cells; liver metabolism; meninges; molecular cloning; neurochemistry; nucleic acid sequence; radionuclides; synaptosomes; tissue /cell culture

The overall goal is to identify the neurochemical mechanisms responsible for ethanol and benzodiazepine intoxication, tolerance and dependence. We postulate that the GABA-activated chloride channel is an important site of action for these drugs and the proposed research will focus exclusively on this receptor-channel complex. Of particular importance is the use of molecular biological techniques to define genetic differences in alcohol and benzodiazepine sensitivity at the level of the individual genes coding for the GABA-activated chloride channel complex. The proposal has 4 aims: 1. GABA-activated chloride channels are modulated by endogenous regulators such as calcium, cAMP and protein kinase C. The possibility that these regulators modulate the acute actions of ethanol or benzodiazepines or are responsible for tolerance to these drugs will be studied. 2. The channel complex consists of alpha, beta and perhaps gamma subunits; these distinct subunits have been cloned and sequenced, but it is not known if they confer different ethanol or benzodiazepine sensitivity on the channel. We will answer this question by testing drug sensitivity after expression of the cloned subunits in oocytes and by creation of transgenic mice with over- or under-expression of genes coding for specific subunits. The transgenic mice will allow us to ascribe functional roles to the different subunits. 3. Brain mRNA from Long/Short-Sleep (LS/SS) and Diazepam Sensitive/Resistant (DS/DR) mice and RNA from cloned genes will be expressed in Xenopus oocytes to allow electrophysiological study of channel function. Cloned genes from drug- sensitive mice (LS,DS) will also be injected into embryos of drug- resistant mice (SS,DR) to produce transgenic mice with expression of genes from resistant and sensitive lines. 4. Genetic differences in behavioral sensitivity to ethanol or benzodiazepines may be due to differences in nucleotide sequences of the genes coding for the subunits of the GABA-activated chloride channel or sequences of genes coding for promoter regions. Use of LS/SS and DS/DR mice for gene cloning will identify sequence differences between the lines.

总体目标是确定负责的神经化学机制 用于乙醇和苯二氮卓类药物中毒，耐受性和依赖性。我们 假设GABA激活的氯化物通道是重要的位置 这些药物的行动和拟议的研究将集中 仅在该受体通道复合物上。特别重要的是 使用分子生物技术来定义遗传差异 在个人水平的酒精和苯二氮卓敏感性中 编码GABA激活的氯化物通道复合物的基因。这 提案有4个目标：1。GABA激活的氯化物通道 由内源性调节剂，例如钙，cAMP和蛋白激酶C。 这些调节器调节乙醇的急性作用的可能性 或苯二氮卓类药物或负责对这些药物的容忍度 被研究。 2。频道建筑群包括Alpha，Beta，也许是 伽玛亚基；这些不同的亚基已被克隆和测序， 但是尚不清楚他们是否赋予不同的乙醇或苯二氮卓 通道上的灵敏度。我们将通过测试药物来回答这个问题 克隆亚基表达卵母细胞和通过 创建具有过度表达基因的转基因小鼠 编码特定亚基。转基因小鼠将使我们能够 将功能角色归因于不同的亚基。 3。来自 长/短囊（LS/SS）和地西epam敏感/抗药性（DS/DR）小鼠 克隆基因的RNA将在爪蟾卵母细胞中表达 通道功能的电生理研究。来自药物的克隆基因 敏感小鼠（LS，DS）也将被注入药物的胚胎 抗性小鼠（SS，DR）产生具有表达的转基因小鼠 来自抗性和敏感线的基因。 4。遗传差异 行为对乙醇或苯二氮卓的敏感性可能是由于 编码亚基的基因的核苷酸序列的差异 GABA激活的氯化物通道或编码的基因序列 发起人区域。将LS/SS和DS/DR小鼠用于基因克隆将 确定线条之间的序列差异。