MITOCHONDRIAL EFFECTS ON SENSITIVITY TO ANESTHETICS

线粒体对麻醉药敏感性的影响

• 批准号: 6343069
• 负责人: Margaret Mary Sedensky
• 金额: $ 25.4万
• 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/6343069
• 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剂的作用原理。这些知识对于未来这些化合物的合理设计无疑至关重要。它也可能被证明是有用的，可以作为理解意识本身基础的探索。我们使用线虫秀丽隐杆线虫来鉴定控制该动物在挥发性麻醉剂中的行为的基因。其中一个基因gas-1的突变对所有测试的挥发性麻醉剂都高度敏感，并且超越任何其他突变对动物在麻醉气体中的行为的影响。 Gas-1 编码线粒体电子传递链第一个蛋白质复合物（复合物 l）的一个亚基。我们建议在分子水平上分析这种蛋白质（NADH 泛醌氧化还原酶的 49kDA 亚基）如何影响线虫的麻醉敏感性。该提案的具体目标是： 1. 在时间和空间上识别该基因的表达。秀丽隐杆线虫独有的强大技术使得了解该基因在其中发挥作用的单个细胞成为可能，从而确定对该基因功能至关重要的发育阶段。 2.测量gas-1对氧化磷酸化和膜过氧化的影响。这两个过程都依赖于复数 l；其中一个或两个可能负责该基因对挥发性麻醉剂行为的控制。 3.发现gas-1和其他线粒体蛋白的其他突变。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gas-1的抑制子或增强子，我们可能会发现编码影响麻醉反应的线粒体蛋白的其他基因。还将研究新鉴定的基因对氧化磷酸化和膜过氧化的影响。我们最终可能会鉴定出一个功能性蛋白质簇，它负责秀丽隐杆线虫中挥发性麻醉剂的作用。 Gas-1 深刻影响线虫的麻醉敏感性；它是该模型中最接近真实麻醉目标的基因。这与早期研究一致，早期研究确定复合物 l 是电子传递链中对挥发性麻醉剂最敏感的蛋白质复合物。准确了解gas-1与挥发性麻醉剂的分子相互作用将为挥发性麻醉剂如何在分子水平发挥作用提供宝贵的信息。