Journal of Applied Physiology AJP: Gastrointestinal and Liver Physiology
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J Appl Physiol 91: 2117-2124, 2001;
8750-7587/01 $5.00
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Vol. 91, Issue 5, 2117-2124, November 2001

Nitric oxide affects sarcoplasmic calcium release in skeletal myotubes

Leo M. A. Heunks1, Herwin A. Machiels1, P. N. Richard Dekhuijzen1, Y. S. Prakash2, and Gary C. Sieck2,3

1 Department of Pulmonary Diseases, University Hospital Nijmegen, Nijmegen, the Netherlands; 2 Departments of Anesthesiology and 3 Physiology and Biophysics, Mayo Clinic and Foundation, Rochester, Minnesota 55905

In the present study, we used real-time confocal microscopy to examine the effects of two nitric oxide (NO) donors on acetylcholine (ACh; 10 µM)- and caffeine (10 mM)-induced intracellular calcium concentration ([Ca2+]i) responses in C2C12 mouse skeletal myotubes. We hypothesized that NO reduces [Ca2+]i in activated skeletal myotubes through oxidation of thiols associated with the sarcoplasmic reticulum Ca2+-release channel. Exposure to diethylamine NONOate (DEA-NO) reversibly increased resting [Ca2+]i level and resulted in a dose-dependent reduction in the amplitude of ACh-induced [Ca2+]i responses (25 ± 7% reduction with 10 µM DEA-NO and 78 ± 14% reduction with 100 µM DEA-NO). These effects of DEA-NO were partly reversible after subsequent exposure to dithiothreitol (10 mM). Preexposure to DEA-NO (1, 10, and 50 µM) also reduced the amplitude of the caffeine-induced [Ca2+]i response. Similar data were obtained by using the chemically distinct NO donor S-nitroso-N-acetyl-penicillamine (100 µM). These results indicate that NO reduces sarcoplasmic reticulum Ca2+ release in skeletal myotubes, probably by a modification of hyperreactive thiols present on the ryanodine receptor channel.

ryanodine receptor; fluorescence; C2C12 myotubes


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