OXYGEN CONCENTRATION & REDOX METABOLISM WITH TRITYL PROBES IN CARDIAC MYOCYTES

氧气浓度

• 批准号: 8364092
• 负责人: JAY Louis ZWEIER
• 金额: $ 0.08万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8364092
• 关键词: Address; Cardiac Myocytes; Cardiomyopathies; Cells; Complement; Esters; Funding; Goals; Grant; Heart; Hypertrophic Cardiomyopathy; Image; Incubated; Life; Location; Measurement; Measures; Metabolism; Microscopy; Muscle Cells; National Center for Research Resources; Oxidation-Reduction; Oxygen; Oxygen saturation measurement; Principal Investigator; Rattus; Research; Research Infrastructure; Resolution; Resources; Source; Spin Labels; Technology; Tissues; United States National Institutes of Health; cellular imaging; cost; extracellular; in vivo; novel

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The goal of this collaborative project is to image O2 and redox metabolism in live cardiac myocytes or myocyte clusters from normal hearts and hearts with cardiomyopathy. Normal cardiac myocytes (from rats) are typically about 20 x 25 x 150 microns. With hypertrophic cardiomyopathy (CM) these cells can increase in size by over 50%. We will make use of recently developed ESRM technology at ACERT and recently developed intra-cellular trityl radical probes and trityl-nitroxide biradicals to provide subcellular and extra cellular imaging of oxygen and redox metabolism that will complement our present "tissue level" imaging on isolated and in vivo hearts, conducted with low field/lower resolution in-vivo ESR imaging. ESR microscopy will be employed with our recently developed novel trityl esters and trityl-nitroxide biradicals that enable measurements of O2 concentrations and redox metabolism in cells and tissues. This is an extension of our grants noted above. Specifically, we will: 1. Incubate and load cardiac myocytes with our recently developed trityl ester probes that we have shown to serve as intracellular spin labels and oximetry probes and then employ ESRM to measure their location and the localized intracellular O2 concentration. Similar measurements will be performed with the extracellular trityl CT-03. 2. Perform similar measurements with our new trityl nitroxide biradicals of intracellular redox metabolism. 3. Subject the cells to different Ca(II) levels and stimulation with beating at rates of 0.5 Hz to 5 Hz and measure the effects on O2 and redox metabolism within the normal and CM myocytes. Several important questions will be addressed including: Is intracellular O2 concentration and redox metabolism different than extracellular values? Are there spatial differences within the cell? Do these values change with myocyte contractile activity? Do these values change with Ca(II) concentration?

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 该合作项目的目标是对正常心脏和心肌病心脏的活心肌细胞或肌细胞簇中的 O2 和氧化还原代谢进行成像。正常心肌细胞（来自大鼠）通常约为 20 x 25 x 150 微米。对于肥厚型心肌病 (CM)，这些细胞的大小会增加 50% 以上。我们将利用 ACERT 最近开发的 ESRM 技术以及最近开发的细胞内三苯甲基自由基探针和三苯甲基氮氧双自由基来提供氧和氧化还原代谢的亚细胞和细胞外成像，这将补充我们目前对分离和氧化还原代谢的“组织水平”成像。体内心脏，采用低场/低分辨率体内 ESR 成像进行。 ESR 显微镜将与我们最近开发的新型三苯甲基酯和三苯甲基氮氧双自由基一起使用，从而能够测量细胞和组织中的 O2 浓度和氧化还原代谢。这是我们上述赠款的延伸。 具体来说，我们将： 1. 用我们最近开发的三苯甲基酯探针孵育并加载心肌细胞，我们已证明这些探针可用作细胞内自旋标签和血氧测定探针，然后使用 ESRM 测量其位置和局部细胞内 O2 浓度。类似的测量将用细胞外三苯甲基 CT-03 进行。 2. 使用我们新的细胞内氧化还原代谢双自由基三苯甲基氮氧自由基进行类似的测量。 3. 将细胞置于不同的 Ca(II) 水平并以 0.5 Hz 至 5 Hz 的频率进行搏动刺激，并测量对正常和 CM 肌细胞内 O2 和氧化还原代谢的影响。 将解决几个重要问题，包括：细胞内 O2 浓度和氧化还原代谢是否与细胞外值不同？细胞内是否存在空间差异？这些值会随着心肌细胞收缩活动而变化吗？这些值会随着 Ca(II) 浓度的变化而变化吗？