Mitochondrial Effects on Sensitivity to Anesthetics

线粒体对麻醉药敏感性的影响

• 批准号: 7652747
• 负责人: Margaret Mary Sedensky
• 金额: $ 8.27万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 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 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7652747
• 关键词: Affect; Anesthesia procedures; Anesthetics; Animal Behavior; Animal Model; Animals; Behavior; Biological Models; Caenorhabditis elegans; Child; Class; Clinical; Complex; Data; Defect; Diagnosis; Free Radicals; Functional disorder; Funding; Gases; Gene Expression; Genes; Homologous Gene; Human; Laboratories; Mammals; Mediating; Metabolic; Metabolism; Microarray Analysis; Mitochondria; Mitochondrial Diseases; Mitochondrial Electron Transport Complex I; Mitochondrial Membrane Protein; Mitochondrial Proteins; Modeling; Molecular; Molecular Profiling; Molecular Target; Mutation; Nematoda; Neurons; Numbers; Oxidative Phosphorylation; Pathway interactions; Patients; Physiological; Point Mutation; Proteins; RNA Interference; Research Personnel; Respiratory physiology; Role; Sorting - Cell Movement; Sum; Testing; Translations; Work; base; behavior change; mutant; novel; programs; response; sevoflurane

DESCRIPTION (provided by applicant): Our laboratory exploits a very simple animal model, the nematode C. elegans, to investigate the molecular mechanism of volatile anesthetic action. We have established that genes and physiologic pathways that alter anesthetic sensitivity in the nematode also alter sensitivity in mammals. The importance of the fact that the same molecular pathways affect sensitivity across different species is hard to overstate. We have identified a group of genes that alter mitochondrial function and control the sensitivity of C. elegans to volatile anesthetics (VAs). A point mutation in gas-1, which encodes the 49 kDa subunit of complex I of the mitochondrial electron transport chain, causes the animal to be very hypersensitive to all VAs. In addition, we have found that a subset of patients, children with metabolic defects most clearly related to complex I function, are profoundly hypersensitive to sevoflurane. This finding provides a clinical correlate to the basic study of very simple animal model. The sum of our work clearly implicates mitochondrial function as a novel mechanism that contributes to the control of anesthetic response. In this proposal we will extend our studies in the nematode to test our hypothesis that alterations in complex I function change anesthetic sensitivity via direct effects on metabolism, and via indirect effects on specific downstream proteins. The specific aims to test these hypotheses are: 1. Using an RNAi screen, determine which subunits of complex I alter anesthetic sensitivity in C. elegans. Our data indicate that the function of this complex is key to mediating behavior in VAs in nematodes, and may have a parallel role in patients with mitochondrial disease. 2. Using gene array analysis, we will determine genes and gene sets whose expression changes as a downstream effect of complex I dysfunction. We will restrict our analysis to expression profiles that alter anesthetic sensitivity by a subtraction strategy that focuses on changes specific to neurons. The microarray analysis will be validated by using RNAi to specifically reduce levels of expression of genes discovered in this aim, and test the resultant effects on behavior in VAs. This superb animal model allows putative molecular targets of VAs to be confirmed or refuted by assessing whole animal behavior. A tractable animal model is crucial to unraveling the mechanism of action of this very important class of anesthetic agents.

描述（由申请人提供）：我们的实验室利用一种非常简单的动物模型，即线虫秀丽隐杆线虫，来研究挥发性麻醉作用的分子机制。我们已经确定，改变线虫麻醉敏感性的基因和生理途径也会改变哺乳动物的敏感性。相同的分子途径影响不同物种的敏感性这一事实的重要性怎么强调都不为过。我们已经确定了一组改变线粒体功能并控制线虫对挥发性麻醉剂 (VA) 敏感性的基因。 Gas-1（编码线粒体电子传递链复合物 I 的 49 kDa 亚基）的点突变导致动物对所有 VA 都非常敏感。此外，我们发现一部分患者，即患有与复合物 I 功能最明显相关的代谢缺陷的儿童，对七氟醚高度过敏。这一发现为非常简单的动物模型的基础研究提供了临床相关性。我们的工作总和清楚地表明线粒体功能是一种有助于控制麻醉反应的新机制。在本提案中，我们将在线虫中扩展我们的研究，以检验我们的假设，即复合体 I 功能的改变通过对代谢的直接影响和对特定下游蛋白质的间接影响来改变麻醉敏感性。检验这些假设的具体目的是： 1. 使用 RNAi 筛选，确定复合物 I 的哪些亚基改变线虫的麻醉敏感性。我们的数据表明，该复合物的功能是介导线虫 VA 行为的关键，并且可能在线粒体疾病患者中发挥平行作用。 2. 使用基因阵列分析，我们将确定其表达随复合体 I 功能障碍的下游效应而变化的基因和基因组。我们将通过关注神经元特异性变化的减法策略将我们的分析限制在改变麻醉敏感性的表达谱上。微阵列分析将通过使用 RNAi 来验证，以特异性降低为此目的发现的基因的表达水平，并测试由此产生的对 VA 行为的影响。这种出色的动物模型可以通过评估整个动物行为来确认或驳斥 VA 的假定分子靶点。易于处理的动物模型对于揭示这一类非常重要的麻醉剂的作用机制至关重要。