Mitochondrial Effects on Sensitivity to Anesthetics

线粒体对麻醉药敏感性的影响

基本信息

批准号：
7930317
负责人：
Margaret Mary Sedensky
金额：
$ 17.14万
依托单位：
SEATTLE CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7930317
关键词：
Affect Anabolism Anesthesia procedures Anesthetics Animal Behavior Animal Model Animals Behavior Behavior Control Binding Binding Sites Caenorhabditis elegans Cells Child Clinical Complex Data Defect Diagnosis Electron Transport Electrons Functional disorder Funding Gases Gene Expression Genes Homologous Gene Human Kinetics Laboratories Location Mammals Measures Mediating Metabolic Metabolism Microarray Analysis Mitochondria Mitochondrial Diseases Mitochondrial Electron Transport Complex I Mitochondrial Proteins Modeling Molecular Molecular Profiling Molecular Target Movement Mutation NADH dehydrogenase (ubiquinone)Nematoda Neurons Oxidative Phosphorylation Pathway interactions Patients Physiological Point Mutation Proteins RNA Interference Research Personnel Respiratory physiology Role Sorting - Cell Movement Sum Testing Translations Ubiquinone Work base behavior change gene discovery mutant novel protein complex response sevoflurane

项目摘要

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.

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，来研究分子挥发性麻醉作用机制。我们已经确定基因和生理途径改变线虫的麻醉敏感性也会改变哺乳动物的敏感性。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并控制线虫对挥发性物质敏感性的基因麻醉剂（VA）。 Gas-1 中的点突变，编码复合物 I 的 49 kDa 亚基线粒体电子传递链，导致动物对所有 VA 都非常敏感。此外，我们发现，患有代谢缺陷的患者子集与复合物 I 最明显相关功能，对七氟烷极度过敏。这一发现提供了与基本的临床相关性研究非常简单的动物模型。我们的工作总和清楚地表明线粒体功能是一种新的功能有助于控制麻醉反应的机制。在本提案中，我们将扩展我们的研究在线虫中检验我们的假设，即复合体 I 功能的改变通过以下途径改变麻醉敏感性：对代谢的直接影响，以及对特定下游蛋白质的间接影响。具体目标是检验这些假设是： 1. 确定复合物 I 的哪些亚基改变线虫的麻醉敏感性。线虫。该复合体的功能是介导线虫 VA 行为的关键，并且可能具有在线粒体疾病患者中具有平行作用。 2. 测量挥发性麻醉剂对泛醌与复合物 I 结合。我们假设该结合位点可能是挥发性化合物的作用目标麻醉剂。 3. 确定其表达随下游效应而变化的基因和基因组复合体 I 功能障碍。我们将仔细地将分析重点放在改变麻醉的表达谱上通过关注神经元特定变化的减法策略来提高敏感性。微阵列分析将通过使用 RNAi 来特异性降低本目标中发现的基因的表达水平进行验证，并测试对 VA 行为的最终影响这种出色的动物模型允许假定的分子目标通过评估整个动物行为来确认或反驳VA。这种易于驯服的动物模型是对于揭示这一类非常重要的麻醉剂的作用机制至关重要。

项目成果

期刊论文数量（14）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Novel interactions between mitochondrial superoxide dismutases and the electron transport chain.

线粒体超氧化物歧化酶和电子传递链之间的新相互作用。

DOI：
发表时间：
2013-12
期刊：
影响因子：
7.8
作者：
Suthammarak, Wichit;Somerlot, Benjamin H;Opheim, Elyce;Sedensky, Margaret;Morgan, Philip G
通讯作者：
Morgan, Philip G

Quality improvement of mitochondrial respiratory chain complex enzyme assays using Caenorhabditis elegans.

使用秀丽隐杆线虫线粒体呼吸链复合酶测定的质量改进。

DOI：
发表时间：
2011-09
期刊：
影响因子：
0
作者：
Chen, Xiulian;Thorburn, David R;Wong, Lee;Vladutiu, Georgirene D;Haas, Richard H;Le, Thuy;Hoppel, Charles;Sedensky, Margaret;Morgan, Philip;Hahn, Si Houn
通讯作者：
Hahn, Si Houn

Propionyl-CoA and adenosylcobalamin metabolism in Caenorhabditis elegans: evidence for a role of methylmalonyl-CoA epimerase in intermediary metabolism.

秀丽隐杆线虫中的丙酰辅酶A和腺苷钴胺代谢：甲基丙二酰辅酶A差向异构酶在中间代谢中作用的证据。

DOI：
10.1016/j.ymgme.2006.06.001
发表时间：
2006-09-01
期刊：
Molecular genetics and metabolism
影响因子：
3.8
作者：
R. Ch;ler;ler;V. Aswani;Matthew S Tsai;Marni J. Falk;Natasha Wehrli;S. Stabler;R. Allen;M. Sedensky
通讯作者：
M. Sedensky

Altered anesthetic sensitivity of mice lacking Ndufs4, a subunit of mitochondrial complex I.

缺乏 Ndufs4（线粒体复合物 I 的一个亚基）的小鼠麻醉敏感性发生改变。