Pflügers Archiv - European Journal of Physiology

, Volume 458, Issue 5, pp 915–928

Reciprocal amplification of ROS and Ca2+ signals in stressed mdx dystrophic skeletal muscle fibers

  • Vyacheslav M. Shkryl
  • Adriano S. Martins
  • Nina D. Ullrich
  • Martha C. Nowycky
  • Ernst Niggli
  • Natalia Shirokova
Muscle Physiology

Abstract

Muscular dystrophies are among the most severe inherited muscle diseases. The genetic defect is a mutation in the gene for dystrophin, a cytoskeletal protein which protects muscle cells from mechanical damage. Mechanical stress, applied as osmotic shock, elicits an abnormal surge of Ca2+ spark-like events in skeletal muscle fibers from dystrophin deficient (mdx) mice. Previous studies suggested a link between changes in the intracellular redox environment and appearance of Ca2+ sparks in normal mammalian skeletal muscle. Here, we tested whether the exaggerated Ca2+ responses in mdx fibers are related to oxidative stress. Localized intracellular and mitochondrial Ca2+ transients, as well as ROS production, were assessed with confocal microscopy. The rate of basal cellular but not mitochondrial ROS generation was significantly higher in mdx cells. This difference was abolished by pre-incubation of mdx fibers with an inhibitor of NAD(P)H oxidase. In addition, immunoblotting showed a significantly stronger expression of NAD(P)H oxidase in mdx muscle, suggesting a major contribution of this enzyme to oxidative stress in mdx fibers. Osmotic shock produced an abnormal and persistent Ca2+ spark activity, which was suppressed by ROS-reducing agents and by inhibitors of NAD(P)H oxidase. These Ca2+ signals resulted in mitochondrial Ca2+ accumulation in mdx fibers and an additional boost in cellular and mitochondrial ROS production. Taken together, our results indicate that the excessive ROS production and the simultaneous activation of abnormal Ca2+ signals amplify each other, finally culminating in a vicious cycle of damaging events, which may contribute to the abnormal stress sensitivity in dystrophic skeletal muscle.

Keywords

Ca2+ sparks Skeletal muscle mdx mice ROS Confocal microscopy 

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Vyacheslav M. Shkryl
    • 1
    • 3
  • Adriano S. Martins
    • 1
  • Nina D. Ullrich
    • 2
  • Martha C. Nowycky
    • 1
  • Ernst Niggli
    • 2
  • Natalia Shirokova
    • 1
  1. 1.Department of Pharmacology and PhysiologyUniversity of Medicine and Dentistry of New JerseyNewarkUSA
  2. 2.Department of PhysiologyUniversity of BernBernSwitzerland
  3. 3.Department of Molecular Biophysics and PhysiologyRush UniversityChicagoUSA

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