MITOCHONDRIAL EFFECTS ON SENSITIVITY TO ANESTHETICS

线粒体对麻醉药敏感性的影响

• 批准号: 6138710
• 负责人: Margaret Mary Sedensky
• 金额: $ 25.82万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6138710
• 关键词: Caenorhabditis elegans; NAD(P)H dehydrogenase; adenosine triphosphate; alternatives to animals in research; anesthetics; cell membrane; drug hypersensitivity; drug screening /evaluation; electron transport; enzyme structure; gene expression; gene mutation; inhalation anesthesia; isoflurane; mitochondria; mutant; nerve /myelin protein; peroxidation; pharmacokinetics; phosphorylation; stereoisomer

The long term goal of this laboratory is to understand at the molecular level how the volatile anesthetics work. Such knowledge is certainly vital for future rational design of these compounds; it may also prove useful as a probe in understanding the basis of consciousness itself. We have used the nematode C. elegans to identify genes that control the behavior of this animal in volatile anesthetics. A mutation in one of these genes, gas-1,is hypersensitive to all volatile anesthetics tested, and overrides the effects of any other mutation on the animal's behavior in anesthetic gases. gas-1 codes for one subunit of the first protein complex (complex l) of the mitochondrial electron transport chain. We propose to analyze at the molecular level how this protein, the 49kDA subunit of NADH ubiquinone oxidoreductase, affects anesthetic sensitivity in C. elegans. The specific aims of this proposal are to: 1. identity the expression of this gene both temporally and spatially. Powerful techniques unique to C. elegans make it possible to know the individual cells in which this gene operates well as to pinpoint developmental stages crucial to the gene's function. 2. measure the effects of gas-1 on oxidative phosphorylation and membrane peroxidation. Both of these processes are dependent on complex l; either or both may be responsible for the gene's control of behavior in volatile anesthetics. 3.find additional mutations in gas-1 and other mitochondrial proteins. We must find more alleles of gas-1 to correlate structure of its protein product to its function. In addition, by screening for suppressors or enhancers of gas-1, we may find other genes that code for mitochondrial proteins that affect anesthetic response. Newly Identified genes will also be studied for their effect on oxidative phosphorylation and membrane peroxidation. We may ultimately identify a functional cluster of proteins that is responsible for the effect of volatile anesthetics in C. elegans. gas-1 profoundly affects anesthetic sensitivity in C. elegans; it is the gene that appears closest to a true anesthetic target in this model. This is consistent with earlier studies which identified complex l as the most sensitive protein complex of the electron transport chain to volatile anesthetics. A precise understanding of gas-1 's molecular interactions with volatile anesthetics will provide invaluable information as to how volatile anesthetics work at the molecular level.

该实验室的长期目标是在分子水平上了解挥发性麻醉药的工作方式。这些知识对于这些化合物的未来理性设计肯定至关重要。它也可能被证明是理解意识本身的基础的调查。我们已经使用线虫C.秀丽隐杆线虫来识别控制该动物在挥发性麻醉剂中行为的基因。这些基因之一的气体-1突变对所有挥发性麻醉剂均过敏，并覆盖了任何其他突变对动物在麻醉气体中行为的影响。气1代码为线粒体电子传输链的第一个蛋白质复合物（复合物L）的一个亚基编码。我们建议在分子水平上分析该蛋白（NADH泛氨基氧化还原酶的49KDA亚基）如何影响秀丽隐杆线虫的麻醉敏感性。该提议的具体目的是：1。身份在时间和空间上表达该基因的表达。秀丽隐杆线虫独有的强大技术使得了解该基因作用的单个细胞，并确定对基因功能至关重要的发育阶段。 2。测量气体1对氧化磷酸化和膜过氧化的影响。这两个过程都取决于复杂的L。无论哪种或两个都可能负责该基因在挥发性麻醉剂中对行为的控制。 3.气体-1和其他线粒体蛋白中的其他突变。我们必须找到更多的GAS-1等位基因，以将其蛋白质产物的结构与其功能相关联。此外，通过筛选抑制器或GAS-1的增强子，我们可能会发现其他基因编码影响麻醉反应的线粒体蛋白。还将研究新鉴定的基因对氧化磷酸化和膜过氧化的影响。我们最终可能会确定蛋白质的功能性簇，该蛋白质负责秀丽隐杆线虫中挥发性麻醉剂的作用。 Gas-1深刻影响秀丽隐杆线虫的麻醉敏感性；在该模型中，它似乎最接近真正的麻醉靶标的基因。这与较早的研究一致，该研究将复合物L视为电子传输链最敏感的蛋白质复合物，以挥发性麻醉。对Gas-1的S分子相互作用与挥发性麻醉药的精确理解将提供有关挥发性麻醉药在分子水平上如何工作的宝贵信息。