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?

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 该合作项目的目标是成像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。正常心肌细胞（来自大鼠）通常为约20 × 25 × 150微米。在肥厚型心肌病（CM）中，这些细胞的大小可以增加50%以上。我们将利用ACERT最近开发的ESRM技术和最近开发的细胞内三苯甲基自由基探针和三苯甲基-氮氧化物双自由基，提供氧和氧化还原代谢的亚细胞和细胞外成像，这将补充我们目前在离体和体内心脏上的“组织水平”成像，采用低场/低分辨率体内ESR成像进行。 ESR显微镜将采用我们最近开发的新的三苯甲基酯和三苯甲基-氮氧双自由基，使测量O2浓度和氧化还原代谢细胞和组织。这是我们上述赠款的延伸。 具体而言，我们将： 1. 用我们最近开发的三苯甲基酯探针孵育和加载心肌细胞，我们已经证明这些探针可用作细胞内自旋标记和血氧测定探针，然后采用ESRM测量它们的位置和局部细胞内O2浓度。将使用细胞外三苯甲基CT-03进行类似测量。 2. 用我们新的细胞内氧化还原代谢的三苯甲基氮氧双自由基进行类似的测量。 3. 使细胞经受不同的Ca（II）水平和以0.5 Hz至5 Hz的速率跳动的刺激，并测量对正常和CM肌细胞内的O2和氧化还原代谢的影响。 几个重要的问题将得到解决，包括：是细胞内的O2浓度和氧化还原代谢比细胞外值不同？细胞内是否存在空间差异？这些值是否随肌细胞收缩活动而变化？这些值随Ca（II）浓度变化吗？