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

  • Muscle Physiology
  • Published:
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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.

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Acknowledgments

This study was supported by the grants from NIH (to N.S.), Muscular Dystrophy Association (to N.S. and M.C.N.), Swiss Foundation for Research on Muscle Diseases (to E.N. and N.S.), Swiss National Science Foundation (to E.N.) and UMDNJ and Sigrist Foundations (to N.S). We thank Dr. Philippe Beauchamp for technical help, and Drs. John Reeves, Roman Shirokov and Andrew Thomas for discussions.

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Author notes

  1. Vyacheslav M. Shkryl

    Present address: Department of Molecular Biophysics and Physiology, Rush University, Chicago, 60612, USA

Authors and Affiliations

  1. Department of Pharmacology and Physiology, University of Medicine and Dentistry of New Jersey, New Jersey Medical School, 185 South Orange Avenue, Newark, NJ, 07103, USA

    Vyacheslav M. Shkryl, Adriano S. Martins, Martha C. Nowycky & Natalia Shirokova

  2. Department of Physiology, University of Bern, Bühlplatz 5, Bern, 3012, Switzerland

    Nina D. Ullrich & Ernst Niggli

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  1. Vyacheslav M. Shkryl
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  2. Adriano S. Martins
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  3. Nina D. Ullrich
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  4. Martha C. Nowycky
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  5. Ernst Niggli
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  6. Natalia Shirokova
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Correspondence to Natalia Shirokova.

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Drs. V.M. Shkryl and A.S. Martins contributed equally to this work.

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Shkryl, V.M., Martins, A.S., Ullrich, N.D. et al. Reciprocal amplification of ROS and Ca2+ signals in stressed mdx dystrophic skeletal muscle fibers. Pflugers Arch - Eur J Physiol 458, 915–928 (2009). https://doi.org/10.1007/s00424-009-0670-2

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