Pflügers Archiv - European Journal of Physiology

, Volume 453, Issue 4, pp 519–529 | Cite as

Contribution from P2X and P2Y purinoreceptors to ATP-evoked changes in intracellular calcium concentration on cultured myotubes

  • Tamás Deli
  • Henrietta Szappanos
  • Gyula Péter Szigeti
  • Julianna Cseri
  • László Kovács
  • László Csernoch
Skeletal Muscle
First Online:
Received:
Accepted:

Abstract

Although the alteration of purinoreceptor pattern on skeletal muscle is known to accompany physiological muscle differentiation and the pathogenesis of muscle dystrophy, the exact identity of and the relative contribution from the individual receptor subtypes to the purinergic signal have been controversial. To identify these subtypes in cultured myotubes of 5–10 nuclei, changes in intracellular calcium concentration and surface membrane ionic currents were detected and calcium fluxes calculated after the application of the subtype-specific agonists 2′3′-O-(benzoyl-4-benzoyl)-ATP (BzATP), 2-methyltio-ADP and UTP. The effectiveness of these agonists together with positive immunocytochemical staining revealed the presence of P2X4, P2X5, P2X7, P2Y1 and P2Y4 receptors. siRNA-reduced protein expression of P2X5, P2X7 and P2Y1 receptors was accompanied by reduction in the ATP-evoked calcium transients. Furthermore, anti-P2X7 siRNA caused a significant drop in the early peak and delayed steady component of the calculated calcium flux. The use of its antagonist, oxidized ATP, similarly to transfection with anti-P2X7 siRNA caused significant reduction in the agonist-elicited ionic currents I ATP and I BzATP, with a greater drop in the latter. Our results demonstrate that the activation of ionotropic P2X4, P2X5 and P2X7 and metabotropic P2Y1 and P2Y4 purinoreceptors participates in forming the calcium transients of multinucleated myotubes.

Keywords

Purinoreceptor subtype Skeletal muscle Intracellular calcium concentration Patch clamp siRNA Calcium flux P2X7 receptor Immunocytochemistry 
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Notes

Acknowledgements

The authors are indebted to G. Czifra for participation in some of the experiments and to R. Öri and I. Vargaand for the excellent technical assistance. This work was supported by OTKA T049151 and OTKA NK61412. G.P. Szigeti holds a Bolyai Fellowship from the Hungarian Academy of Sciences.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Tamás Deli
    • 1
  • Henrietta Szappanos
    • 1
  • Gyula Péter Szigeti
    • 1
  • Julianna Cseri
    • 1
  • László Kovács
    • 1
    • 2
  • László Csernoch
    • 1
    • 2
  1. 1.Department of Physiology, Medical and Health Science CentreUniversity of DebrecenDebrecenHungary
  2. 2.Cell Physiology Research Group of the Hungarian Academy of Sciences, Research Centre for Molecular MedicineMedical and Health Science Centre, University of DebrecenDebrecenHungary

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