Nitric oxide affects sarcoplasmic calcium release in skeletal myotubes

Leo M A Heunks, Herwin A. Machiels, P. N Richard Dekhuijzen, Y. S. Prakash, Gary C. Sieck

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

In the present study, we used real-time confocal microscopy to examine the effects of two nitric oxide (NO) donors on acetyl-choline (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.

Original languageEnglish
Pages (from-to)2117-2124
Number of pages8
JournalJournal of Applied Physiology
Volume91
Issue number5
Publication statusPublished - 10 Nov 2001

Cite this

Heunks, L. M. A., Machiels, H. A., Dekhuijzen, P. N. R., Prakash, Y. S., & Sieck, G. C. (2001). Nitric oxide affects sarcoplasmic calcium release in skeletal myotubes. Journal of Applied Physiology, 91(5), 2117-2124.
Heunks, Leo M A ; Machiels, Herwin A. ; Dekhuijzen, P. N Richard ; Prakash, Y. S. ; Sieck, Gary C. / Nitric oxide affects sarcoplasmic calcium release in skeletal myotubes. In: Journal of Applied Physiology. 2001 ; Vol. 91, No. 5. pp. 2117-2124.
@article{e50f86876a6e4bf98bddcf1d4a2f5560,
title = "Nitric oxide affects sarcoplasmic calcium release in skeletal myotubes",
abstract = "In the present study, we used real-time confocal microscopy to examine the effects of two nitric oxide (NO) donors on acetyl-choline (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.",
keywords = "CC myotubes, Fluorescence, Ryanodine receptor",
author = "Heunks, {Leo M A} and Machiels, {Herwin A.} and Dekhuijzen, {P. N Richard} and Prakash, {Y. S.} and Sieck, {Gary C.}",
year = "2001",
month = "11",
day = "10",
language = "English",
volume = "91",
pages = "2117--2124",
journal = "Journal of Applied Physiology",
issn = "8750-7587",
publisher = "American Physiological Society",
number = "5",

}

Heunks, LMA, Machiels, HA, Dekhuijzen, PNR, Prakash, YS & Sieck, GC 2001, 'Nitric oxide affects sarcoplasmic calcium release in skeletal myotubes' Journal of Applied Physiology, vol. 91, no. 5, pp. 2117-2124.

Nitric oxide affects sarcoplasmic calcium release in skeletal myotubes. / Heunks, Leo M A; Machiels, Herwin A.; Dekhuijzen, P. N Richard; Prakash, Y. S.; Sieck, Gary C.

In: Journal of Applied Physiology, Vol. 91, No. 5, 10.11.2001, p. 2117-2124.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Nitric oxide affects sarcoplasmic calcium release in skeletal myotubes

AU - Heunks, Leo M A

AU - Machiels, Herwin A.

AU - Dekhuijzen, P. N Richard

AU - Prakash, Y. S.

AU - Sieck, Gary C.

PY - 2001/11/10

Y1 - 2001/11/10

N2 - In the present study, we used real-time confocal microscopy to examine the effects of two nitric oxide (NO) donors on acetyl-choline (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.

AB - In the present study, we used real-time confocal microscopy to examine the effects of two nitric oxide (NO) donors on acetyl-choline (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.

KW - CC myotubes

KW - Fluorescence

KW - Ryanodine receptor

UR - http://www.scopus.com/inward/record.url?scp=0034752059&partnerID=8YFLogxK

M3 - Article

VL - 91

SP - 2117

EP - 2124

JO - Journal of Applied Physiology

JF - Journal of Applied Physiology

SN - 8750-7587

IS - 5

ER -

Heunks LMA, Machiels HA, Dekhuijzen PNR, Prakash YS, Sieck GC. Nitric oxide affects sarcoplasmic calcium release in skeletal myotubes. Journal of Applied Physiology. 2001 Nov 10;91(5):2117-2124.