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Integrin stimulation induces calcium signalling in rat cardiomyocytes by a NO-dependent mechanism

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Abstract

The myocardial stretch-induced increase in intracellular [Ca2+] ([Ca2+]i) is considered to be caused by integrin stimulation. Myocardial stretch is also associated with increased nitric oxide (NO) formation. We hypothesised that NO is implicated in calcium signalling following integrin stimulation. Integrins of neonatal rat cardiomyocytes were stimulated with a pentapeptide containing the Arg-Gly-Asp (RGD) sequence. [Ca2+]i was measured with Fura2, [NO]i was measured with DAF2 and phosphorylation of focal adhesion kinase (FAK) was monitored with immunofluorescence techniques. Integrin stimulation increased both [NO]i and [Ca2+]i, the latter response being inhibited by ryanodine receptor-2 (RyR2) blockers and by NG-monomethyl-L-arginine (L-NMMA), an inhibitor of NOS, but resistant to GdCl3, diltiazem and wortmannin. Integrin-induced intracellular Ca2+ release thus appears to be independent of the influx of extracellular Ca2+ and phosphatidylinositol-3 kinase activity. In addition, integrin stimulation induced phosphorylation of FAK. Our results provide evidence for an integrin-induced Ca2+ release from RyR2 which is mediated by NO formation, probably via FAK-induced NOS activation.

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Abbreviations

DAF2-DA:

4,5-diaminofluorescein diacetate

FAK:

Focal adhesion kinase

L-NMMA:

NG-monomethyl-L-arginine

NO:

Nitric oxide

NOS:

Nitric oxide synthase

PI3K:

Phosphatidylinositol-3 kinase

P+RR:

Procaine plus ruthenium red

RGD:

Arg-Gly-Asp sequence

RGE:

Asp-Gly-Arg sequence

RyR:

Ryanodine receptor

SNAP:

S-nitroso-N-acetylpenicillamine

SNP:

Sodium-nitroprusside

SR:

Sarcoplasmic reticulum

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Acknowledgements

This study was supported by the Dutch Heart Foundation #97.176. We thank Salma Gurmany for assistance with NO imaging.

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Authors and Affiliations

  1. Department of Cardiology, Leiden University Medical Centre, P.O. Box 9600, 2300, RC, Leiden, The Netherlands

    C. G. C. van der Wees, W. H. Bax, E. J. M. van der Valk & A. van der Laarse

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  1. C. G. C. van der Wees
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  2. W. H. Bax
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  3. E. J. M. van der Valk
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  4. A. van der Laarse
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Correspondence to A. van der Laarse.

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van der Wees, C.G.C., Bax, W.H., van der Valk, E.J.M. et al. Integrin stimulation induces calcium signalling in rat cardiomyocytes by a NO-dependent mechanism. Pflugers Arch - Eur J Physiol 451, 588–595 (2006). https://doi.org/10.1007/s00424-005-1402-x

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  • DOI: https://doi.org/10.1007/s00424-005-1402-x

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