Simple Quantitative Detection of Mitochondrial Superoxide Production in Live Cells.
From: Section on Oxidative Stress Tissue Injury, Laboratory of Physiological Studies, National Institutes of Health/NIAAA, Bethesda, MD 20892-9413, USA.
Biochemical and biophysical research communications
- Publish Date: Jun 2007
- ISSN: 0006-291X
- Volume: 358
- Issue: 1
- Pages: 203-8
- Medium: Print
- Language: English
- Citation (JAMA): Mukhopadhyay Partha, Rajesh Mohanraj, Yoshihiro Kashiwaya, et al. Simple Quantitative Detection of Mitochondrial Superoxide Production in Live Cells.. Biochem. Biophys. Res. Commun. Jun 2007;358:203-8
Abstract
Experiments with isolated mitochondria have established that these organelles are pivotal intracellular sources of superoxide in a variety of pathophysiological conditions. Recently, a novel fluoroprobe MitoSOX Red was introduced for selective detection of superoxide in the mitochondria of live cells and was validated with confocal microscopy. Here we show approximately 3-7 fold dose- and time-dependent increase in mitochondrial superoxide production (measured by MitoSOX using flow cytometry and confocal microscopy) in rat cardiac derived H9c2 myocytes and/or in human coronary artery endothelial cells triggered by Antimycin A, Paraquat, Doxorubicin or high glucose. These results establish a novel, quantitative method for simple detection of mitochondrial superoxide generation simultaneously in a large population of live cells by flow cytometry. This method can also be adapted for immune cell studies with mixed population of T or B cells or their subsets to analyze mitochondrial superoxide levels using multiple labeled surface markers in individual populations.
Mesh Headings (Keywords): Animals, Antimycin A, Cells, Cultured, Coronary Vessels, Doxorubicin, Endothelial Cells, Endothelium, Vascular, Flow Cytometry, Fluorescent Dyes, Glucose, Humans, Microscopy, Confocal, Mitochondria, Heart, Myocytes, Cardiac, Paraquat, Phenanthridines, Rats, Superoxides
Check for Full Text / PubMed Unique Identifier (PMID): 17475217
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