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

被引:21
作者
Marsigliante, S
Elia, MG
Di Jeso, B
Greco, S
Muscella, A
Storelli, C
机构
[1] Univ Lecce, Dept Biol, Physiol Lab, I-73100 Lecce, Italy
[2] Univ Lecce, Dept Biol, Lab Gen Pathol, I-73100 Lecce, Italy
关键词
PC-Cl3; thyroid; ATP; P2Y; Ca2+;
D O I
10.1016/S0898-6568(01)00208-X
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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 InsP(3) 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 muM 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. (C) 2002 Elsevier Science Inc. All rights reserved.
引用
收藏
页码:61 / 67
页数:7
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