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.

我们的实验室利用了一个非常简单的动物模型，线虫C.秀拉斯绿杆菌，以研究分子挥发性麻醉作用的机制。我们已经确定了基因和生理途径在线虫中改变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灵敏度很难夸大其词。我们有鉴定出一组改变线粒体功能并控制秀丽隐杆线虫对挥发性的敏感性的基因麻醉药（VAS）。 GAS-1中的点突变，该突变编码了49 kDa的复合物I 线粒体电子传输链使动物对所有VA都非常敏感。此外，我们发现，一部分患者，代谢缺陷的儿童与复合物I最明显有关功能对Sevoflurane非常高度敏感。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。我们工作的总和清楚地暗示了线粒体功能作为一种新颖有助于控制麻醉反应的机制。在此提案中，我们将扩展我们的研究在测试我们的假设的线虫中，复杂I功能的改变通过直接对代谢以及通过间接影响对特定下游蛋白的影响。具体目的测试这些假设是：1。确定复合物I的哪些亚基改变了C中的麻醉敏感性。秀丽隐杆线。该复合物的功能是介导线虫中VA的行为的关键，并且可能具有线粒体疾病患者的平行作用。 2。测量挥发性麻醉药对泛氨基酮与复合物I的结合。我们假设该结合位点可能是挥发性的作用目标麻醉药。 3。确定基因和基因集，其表达随着下游效应而变化复杂的I功能障碍。我们将仔细将分析集中于改变麻醉剂的表达曲线通过减法策略的敏感性，该策略的重点是针对神经元的变化。微阵列分析将通过使用RNAi专门降低Thisaim中发现的基因表达水平来验证并测试对VAS中行为的影响的影响，这种出色的动物模型允许假定的分子靶标通过评估全动物行为可以确认或驳斥的VA。这样的可拖动动物模型是对于阐明这种非常重要的麻醉剂的作用机理至关重要。

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）

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

DOI：
10.1371/journal.pone.0042904
发表时间：
2012
期刊：
PloS one
影响因子：
3.7
作者：
Quintana A;Morgan PG;Kruse SE;Palmiter RD;Sedensky MM
通讯作者：
Sedensky MM

Subcomplex Ilambda specifically controls integrated mitochondrial functions in Caenorhabditis elegans.

DOI：
10.1371/journal.pone.0006607
发表时间：
2009-08-12
期刊：
PloS one
影响因子：
3.7
作者：
Falk MJ;Rosenjack JR;Polyak E;Suthammarak W;Chen Z;Morgan PG;Sedensky MM
通讯作者：
Sedensky MM

Comparison of proteomic and metabolomic profiles of mutants of the mitochondrial respiratory chain in Caenorhabditis elegans.

DOI：
10.1016/j.mito.2014.12.004
发表时间：
2015-01
期刊：
MITOCHONDRION
影响因子：
4.4
作者：
Morgan, P. G.;Higdon, R.;Kolker, N.;Bauman, A. T.;Ilkayeva, O.;Newgard, C. B.;Kolker, E.;Steele, L. M.;Sedensky, M. M.
通讯作者：
Sedensky, M. M.

Altered redox status of coenzyme Q9 reflects mitochondrial electron transport chain deficiencies in Caenorhabditis elegans.

辅酶 Q9 氧化还原状态的改变反映了秀丽隐杆线虫线粒体电子传递链的缺陷。