Increase of [Ca2+]i via activation of ATP receptors in PC-Cl3 rat thyroid cell line

Santo Marsigliante, Maria Giovanna Elia, Bruno Di Jeso, Simona Greco, Antonella Muscella, Carlo Storelli

Research output: Contribution to journalArticlepeer-review

Abstract

In PC-Cl3 rat thyroid cell line, ATP and UTP provoked a transient increase in [Ca2+]i, followed by a lower sustained phase. Removal of extracellular Ca2+ reduced the initial transient response and completely abolished the plateau phase. Thapsigargin (TG) caused a rapid rise in [Ca2+]i and subsequent addition of ATP was without effect. The transitory activation of [Ca2+]i was dose-dependently attenuated in cells pretreated with the specific inhibitor of phospholipase C (PLC), U73122. These data suggest that the ATP-stimulated increment of [Ca2+]i required InsP3 formation and binding to its specific receptors in Ca2+ stores. Desensitisation was demonstrated with respect to the calcium response to ATP and UTP in Fura 2-loaded cells. Further studies were performed to investigate whether the effect of ATP on Ca2+ entry into PC-Cl3 cells was via L-type voltage-dependent Ca2+ channels (L-VDCC) and/or by the capacitative pathway. Nifedipine decreased ATP-induced increase on [Ca2+]i. Addition of 2 mM Ca2+ induced a [Ca2+]i rise after pretreatment of the cells with TG or with 100 μM ATP in Ca2+-free medium. These data indicate that Ca2+ entry into PC-Cl3 stimulated with ATP occurs through both an L-VDCC and through a capacitative pathway. Using buffers with differing Na+ concentrations, we found that the effects of ATP were dependent of extracellular Na+, suggesting that a Na+/Ca2+ exchange mechanism is also operative. These data suggest the existence, in PC-Cl3 cell line, of a P2Y purinergic receptor able to increase the [Ca2+]i via PLC activation, Ca2+ store depletion, capacitative Ca2+ entry and L-VDCC activation.

Original languageEnglish
Pages (from-to)61-67
Number of pages7
JournalCellular Signalling
Volume14
Issue number1
DOIs
Publication statusPublished - 2002

Keywords

  • ATP
  • Ca
  • P2Y
  • PC-Cl3
  • Thyroid

ASJC Scopus subject areas

  • Cell Biology

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