Simple quantitative detection of mitochondrial superoxide production in live cells

Partha Mukhopadhyay, Mohanraj Rajesh, Kashiwaya Yoshihiro, G. Haskó, Pál Pacher

Research output: Contribution to journalArticle

216 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

Fingerprint

Superoxides
Mitochondria
Flow cytometry
Confocal microscopy
Confocal Microscopy
Flow Cytometry
Population
Antimycin A
Paraquat
Endothelial cells
Organelles
Doxorubicin
Muscle Cells
Rats
Coronary Vessels
B-Lymphocytes
Endothelial Cells
Cells
T-Lymphocytes
Glucose

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Simple quantitative detection of mitochondrial superoxide production in live cells. / Mukhopadhyay, Partha; Rajesh, Mohanraj; Yoshihiro, Kashiwaya; Haskó, G.; Pacher, Pál.

In: Biochemical and Biophysical Research Communications, Vol. 358, No. 1, 22.06.2007, p. 203-208.

Research output: Contribution to journalArticle

Mukhopadhyay, Partha ; Rajesh, Mohanraj ; Yoshihiro, Kashiwaya ; Haskó, G. ; Pacher, Pál. / Simple quantitative detection of mitochondrial superoxide production in live cells. In: Biochemical and Biophysical Research Communications. 2007 ; Vol. 358, No. 1. pp. 203-208.
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