Mitochondrial Effects on Sensitivity to Anesthetics

我们的实验室利用一个非常简单的动物模型，线虫线虫，来研究分子 挥发性麻醉剂的作用机制。我们已经确定了基因和生理途径 改变线虫的麻醉敏感度也会改变哺乳动物的敏感度。这个事实的重要性 相同的分子途径影响不同物种的敏感性，这一点怎么强调都不为过。我们有 确定了一组改变线粒体功能和控制线虫对挥发性物质敏感性的基因 麻醉药(VAS)。GAS-1中的一个点突变，它编码的是该病毒复合体I的49 kDa亚基 线粒体电子传输链，导致动物对所有VAS非常敏感。此外, 我们已经发现，有代谢缺陷的儿童患者的一部分最明显地与复合体I有关 功能，对七氟醚极度敏感。这一发现为临床提供了与基本的 研究非常简单的动物模型。我们的工作总结清楚地表明线粒体的功能是一种新的 有助于控制麻醉反应的机制。在这项建议中，我们将延长我们的研究 在线虫中测试我们的假设，即复合体I功能的变化通过 对新陈代谢的直接影响，以及对特定下游蛋白质的间接影响。具体目的是 检验这些假设是：1.确定复合体的哪些亚基改变了C。 优雅女装。这个复合体的功能是调节线虫VAS行为的关键，并可能具有 在线粒体疾病患者中的平行作用。2.测量挥发性麻醉药对患者的影响 泛醌与络合物的结合。我们推测这个结合部位可能是挥发性物质的作用靶点。 麻醉剂。3.确定其表达变化作为下游效应的基因和基因集 复杂性I型功能障碍。我们将仔细分析改变麻醉剂的表达模式 通过专注于神经元特有变化的减法策略提高敏感度。基因芯片分析 将通过使用RNAi来具体降低在这一目标中发现的基因的表达水平来进行验证， 并在VAS中测试结果对行为的影响这种极好的动物模型允许假定的分子靶标 通过评估整个动物的行为来证实或驳斥VAS的观点。这样一个容易驯服的动物模型是 对于解开这类非常重要的麻醉剂的作用机制至关重要。

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. Determine which subunits of complex I alter anesthetic sensitivity in C. elegans. 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. Measure the effect of volatile anesthetics on ubiquinone binding to complex I. We hypothesize that this binding site may be a target of action for volatile anesthetics. 3. Determine genes and gene sets whose expression changes as a downstream effect of complex I dysfunction. We will carefully focus 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 thisaim, 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. Such a tractable animal model is crucial to unraveling the mechanism of action of this very important class of anesthetic agents.