Simple quantitative detection of mitochondrial superoxide production in live cells

Partha Mukhopadhyay, Mohanraj Rajesh, Kashiwaya Yoshihiro, György Haskó, Pál Pacher

Research output: Contribution to journalArticlepeer-review

255 Citations (Scopus)

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 ∼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.

Original languageEnglish
Pages (from-to)203-208
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume358
Issue number1
DOIs
Publication statusPublished - Jun 22 2007
Externally publishedYes

Keywords

  • Antimycin A (AntA)
  • Cardiomyocytes
  • Doxorubicin (DOX)
  • Free radicals
  • H9c2
  • Human coronary artery endothelial cells (HCAECs)
  • MitoSOX
  • Mitochondria
  • Paraquat (PQ)
  • Superoxide

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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