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?

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 这个协作项目的目的是在现场心肌细胞或心脏肌病心脏的心脏肌细胞或心脏症状的心肌簇中对O2和氧化还原代谢进行图像。正常心肌细胞（来自大鼠）通常约为20 x 25 x 150微米。使用肥厚的心肌病（CM），这些细胞的大小可以增加50％以上。 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显微镜将用于我们最近开发的新型Trityl酯和三乙二醇Biradicals，可实现细胞和组织中O2浓度和氧化还原代谢的测量。这是上面指出的我们的赠款的延伸。 具体来说，我们将： 1。与我们最近开发的Trityl酯探针一起孵育和负载心肌细胞，我们已证明它们可以用作细胞内的自旋标记和血氧仪探针，然后采用ESRM来测量其位置和局部的细胞内O2浓度。将使用细胞外Trityl CT-03进行类似的测量。 2。对我们的新三元二氧化物生物自由基对细胞内氧化还原代谢进行类似的测量。 3。使细胞以0.5 Hz至5 Hz的速度进行不同的Ca（II）水平和刺激，并测量正常和CM肌细胞内对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不同于细胞外值吗？细胞内有空间差异吗？这些值会随着心肌收缩活动而变化吗？这些值会随CA（ii）浓度而变化吗？