Mapping of redox state of mitochondrial cytochromes in live cardiomyocytes using Raman microspectroscopy

PLoS One. 2012;7(9):e41990. doi: 10.1371/journal.pone.0041990. Epub 2012 Sep 5.

Abstract

This paper presents a nonivasive approach to study redox state of reduced cytochromes c, c1 and b of complexes II and III in mitochondria of live cardiomyocytes by means of Raman microspectroscopy. For the first time with the proposed approach we perform studies of rod- and round-shaped cardiomyocytes, representing different morphological and functional states. Raman mapping and cluster analysis reveal that these cardiomyocytes differ in the amounts of reduced cytochromes c, c1 and b. The rod-shaped cardiomyocytes possess uneven distribution of reduced cytochromes c, c1 and b in cell center and periphery. Moreover, by means of Raman spectroscopy we demonstrated the decrease in the relative amounts of reduced cytochromes c, c1 and b in the rod-shaped cardiomyocytes caused by H2O2-induced oxidative stress before any visible changes. Results of Raman mapping and time-dependent study of reduced cytochromes of complexes II and III and cytochrome c in cardiomyocytes are in a good agreement with our fluorescence indicator studies and other published data.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cytochromes / metabolism*
  • Hydrogen Peroxide / chemistry
  • Hydroxyl Radical
  • Membrane Potentials
  • Microscopy, Fluorescence / methods
  • Mitochondria / metabolism*
  • Models, Statistical
  • Myocytes, Cardiac / cytology
  • Oxidation-Reduction*
  • Oxidative Stress
  • Rats
  • Spectrum Analysis, Raman / methods*
  • Time Factors

Substances

  • Cytochromes
  • Hydroxyl Radical
  • Hydrogen Peroxide

Grant support

Support for this work came from The Danish Council for Independent Research | Natural Sciences (to N.A.B. and O.V.S.). M.T and N.L.F. were supported by The Danish National Research Foundation for Heart Arrhythmia, The Novo Nordisk Foundation and the A. and E. Danielsen's Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.