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.

这个实验室的长期目标是在分子水平上了解挥发性麻醉剂是如何发挥作用的。这样的知识对于这些化合物未来的合理设计当然是至关重要的；它也可能被证明是理解意识本身基础的一个有用的探测器。我们已经使用线虫线虫来识别在挥发性麻醉剂中控制这种动物行为的基因。其中一个基因的突变，GAS-1，对所有测试的挥发性麻醉剂都是超敏的，并超过了任何其他突变对动物在麻醉气体中行为的影响。GAS-1编码线粒体电子传递链第一个蛋白质复合体(L复合体)的一个亚基。我们建议在分子水平上分析这种NADH泛醌氧化还原酶49kDA亚基蛋白如何影响线虫的麻醉敏感性。这一建议的具体目的是：1.在时间和空间上鉴定该基因的表达。线虫特有的强大技术使人们有可能了解这种基因在其中很好地发挥作用的单个细胞，并精确定位对该基因功能至关重要的发育阶段。2.检测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杂的L；其中一个或两个都可能是该基因控制挥发性麻醉剂行为的原因。3.在GAS-1和其他线粒体蛋白中发现额外的突变。我们必须找到更多的GAS-1等位基因，才能将其蛋白产物的结构与其功能联系起来。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。还将研究新发现的基因对氧化磷酸化和膜过氧化的影响。我们最终可能确定一个蛋白质的功能簇，负责挥发性麻醉剂对线虫的影响。GAS-1深刻地影响线虫的麻醉敏感性；在这个模型中，它是最接近真正的麻醉靶点的基因。这与早期的研究一致，即L复合体是电子传递链上对挥发性麻醉药最敏感的蛋白质复合体。准确了解GAS-1‘S与挥发性麻醉药的分子相互作用将为了解挥发性麻醉剂在分子水平上的作用机制提供宝贵的信息。