Mechanism of ATP-induced [Ca2+]i mobilization in rat basilar smooth muscle cells

被引:16
作者
Aoki, K [1 ]
Zubkov, AY [1 ]
Parent, AD [1 ]
Zhang, JH [1 ]
机构
[1] Univ Mississippi, Med Ctr, Dept Neurosurg, Jackson, MS 39216 USA
关键词
calcium; protein kinases; protein-tyrosine kinase; receptors; purinergic P-2; rats;
D O I
10.1161/01.STR.31.6.1377
中图分类号
R74 [神经病学与精神病学];
学科分类号
摘要
Background and Purpose-We have previously reported that extracellular ATP activates P-2u receptors and increases intracellular free Ca2+ ([Ca2+](i)) by G protein/phospholipase C/inositol 1,4,5-triphosphate pathways in cerebral artery smooth muscle cells. However, the possible contribution of other signaling pathways remains unclear. This study was undertaken to investigate the role of protein tyrosine kinase (PTK) and mitogen-activated protein kinase (MAPK) in mediating ATP-induced Ca2+ mobilization in rat basilar artery smooth muscle cells (RBASMCs). Methods-RBASMCs were freshly isolated, and [Ca2+](i) was monitored by fura 2 microfluorimetry. MAPK phosphorylation was studied by the Western blot technique. Results-ATP produced a biphasic [Ca2+](i) response, which consists of releasing Ca2+ from internal stores and influx from extracellular space. PTK inhibitors tyrphostin 51 and genistein inhibited [Ca2+](i) response to ATP. Tyrphostin Al, an inactive analogue of tyrphostins, failed to reduce the ATP-induced response. MAPK kinase inhibitor PD98059, but not U0126, reduced the ATP-induced [Ca2+](i) response. Phosphatidylinositol 3-kinase (PI3-K) tyrosine kinase inhibitor wortmannin, but not janus tyrosine kinase (JAK2) inhibitor AG490, partially inhibited the [Ca2+], response induced by ATP. In addition, ATP enhanced MAPK phosphorylation in a concentration- and time-dependent manner, and genistein, tyrphostin 51, PD98059, and U0126 inhibited MAPK phosphorylation. Conclusions-Extracellular ATP produced [Ca2+](i) elevation and MAPK phosphorylation in RBASMCs, and the effect was regulated by PTK. The role of MAPK in ATP-induced [Ca2+](i) elevation is not clear. PI3-K tyrosine kinase and JAK2 tyrosine kinase may not play an important role in the ATP-induced [Ca2+](i) response in RBASMCs.
引用
收藏
页码:1377 / 1384
页数:8
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