Mitochondria/Mass Spectroscopy Core

线粒体/质谱核心

• 批准号: 7750211
• 负责人: Charles Leslie Hoppel
• 金额: $ 28.67万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-14 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7750211
• 关键词: Affect; Attention; Canis familiaris; Cardiac; Cardiolipins; Cells; Complex; Electron Transport; Electrophoresis; Energy Metabolism; Functional disorder; Heart; Heart failure; Individual; Instruction; Isoprostanes; Lipids; Malonyl Coenzyme A; Mass Spectrum Analysis; Measures; Membrane; Mitochondria; Modification; Mus; Muscle Cells; Myocardium; Myofibrils; Oxidative Stress; Phospholipids; Physiological; Population; Program Research Project Grants; Quality Control; Sampling; Services; Skin; Tissues; Transmission Electron Microscopy; Trypsin; acylcarnitine; analytical method; complex IV; in vivo; oxidation; oxidized low density lipoprotein; research study

Cardiac mitochondria exist in two distinct populations wathin the myocyte, subsarcolemmal mitochondria (SSM), located beneath the sarcolemmal membrane, and interfibrillar mitochondria (IFM) that reside between the myofibrils. SSM are released by tissue homogenization, whereas the liberation of IFM requires a brief exposure of the resulting skinned myocytes to trypsin treatment. The two populations of mitochondria are differently affected under both physiological and pathophysiological conditions and thus requires to study of each individual population separately. The Mitochondrial/Mass Spec Core (Core D) will perform the following experiments as a service to the different Projects. 1. Isolate subsarcolemmal and interfibrillar mitochondria from experimental hearts; assess quality control. 2. Obtain samples of myocardium and isolated populations of SSM and IFM for electron transport chain complex analysis. 3. Obtain samples of heart, SSM, and IFM for lipid extraction and phospholipid analysis. Special attention is paid to cardiolipin (CL) analysis: CL content, CL species and CL oxidation. 4. Aquire heart tissue for analysis of acyl-CoAs and acylcarnitines with special emphasis on malonyl-CoA a key metabolite in switch in fuel utilization. 5. Carry out mass spectrometric analysis to identify posttranslational and oxidative modifications on ETC subunits, especially complex IV. 6. Perform structural analysis on cardiac tissue and cardiac SSM and IFM by transmission electron microscopy. 7. Isoprostane and oxidized low density lipoprotein (oxLDL) quantitation as a measure of in vivo oxidative stress. 8. Blue Native Electrophoresis will be done on mitochondria from Project 1 and 4. RELEVANCE (See instructions): Mitochondrial are the powerhouses and the cell. During heart failure an energy crises occurs in the heart and is related to the dysfunction of mitochondria. This Core will provide mitochondrial isolation from heart of dogs and mice under experimental conditions and perform analytical methods to support the 4 project within the Program Project Grant.

心脏线粒体存在于两个不同种群中，肌细胞肌细胞，亚囊膜线粒体（SSM）， 位于肌膜膜下方，以及位于岩膜之间 肌原纤维。 SSM通过组织均质化释放，而IFM的解放需要短暂的暴露 由此产生的皮肤肌细胞进行胰蛋白酶治疗。线粒体的两个种群有所不同 在生理和病理生理条件下受到影响，因此需要研究每一种 单独人口。线粒体/质量规格核心（核心D）将执行以下 实验作为对不同项目的服务。 1。分离的肾小管和纤维间线粒体的实验心脏；评估质量控制。 2。获得电子传输链复合物的心肌和IFM的孤立人群的样品 分析。 3。获取心脏，SSM和IFM的样品进行脂质提取和磷脂分析。特别注意 到Cardiolipin（CL）分析：CL含量，CL物种和CL氧化。 4。用于分析酰基-COAS和酰基肉碱的Aquire心脏组织，特别强调丙酰coA 燃料利用中的开关中的代谢物。 5。进行质谱分析，以确定翻译后和氧化修饰等 亚基，特别是复杂的IV。 6。通过透射电子显微镜对心脏组织，心脏SSM和IFM进行结构分析。 7。异丙烷和氧化的低密度脂蛋白（OXLDL）定量，作为体内氧化的量度 压力。 8。蓝色天然电泳将从项目1和4的线粒体上完成。 相关性（请参阅说明）： 线粒体是强力室和细胞。在心力衰竭期间，能量危机发生在心脏中 与线粒体的功能障碍有关。该核心将提供从心脏的线粒体隔离 狗和小鼠在实验条件下并执行分析方法以支持4个项目 计划项目赠款。