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Título: | Calcium signalling through nucleotide receptor P(2Y2) in cultured human vascular endothelium |
Autor: | Viana, Félix CSIC ORCID ; Smedt, Humbert de; Droogmans, Guy; Nilius, Bernd | Fecha de publicación: | 1998 | Editor: | Elsevier | Citación: | Cell Calcium 24(2): 117-127 (1998) | Resumen: | Microfluorometric measurements in Fura-2-loaded single cultured human vascular endothelial cells were used to characterize the intracellular calcium [Ca2+]i responses triggered by extracellular application of adenosine 5′-triphosphate (ATP) and other nucleotides. Application of ATP or uridine 5′-triphosphate (UTP) gave rise to dose-dependent elevations of [Ca22+]i in all the cells tested. At saturating concentrations of agonist, the [Ca2+]i response was biphasic, with an early peak and a sustained plateau. Unlike peak responses, the sustained Ca2+ plateau was sensitive to removal of Ca2+ from the external medium. Mn2+ quenching revealed the presence of Ca2+ influx during the agonist-induced calcium plateau. The agonist-evoked calcium plateau was inhibited in a dose-dependent manner by the Cl-channel blocker NPPB, by the divalent cation Ni2+ and by the imidazole antimycotic econazole. Previously, these compounds have been shown to block store-operated Ca2+ entry. The two phases of the agonist-evoked [Ca2+]i response were blocked by the specific phospholipase C inhibitor U-73122 and by intracellular injection of low molecular weight heparin, suggesting the involvement of IP3 sensitive intracellular Ca2+ stores. The pharmacological profile of the response, using different nucleotides and analogues, ATP = UTP > ADP = UDP, and no responses to P2x1 and P2Y1 agonists, suggested the involvement of P2Y2 receptors. The expression of mRNA for the P2Y2 receptor was detected by RT-PCR analysis. These results indicate that P2Y2 receptors linked to intracellular Ca2+ mobilization are present in human vascular endothelial cells. The initial [Ca2+]i mobilization is followed by a phase of elevated [Ca2+]i influx. | Versión del editor: | https://doi.org/10.1016/S0143-4160(98)90079-3 | URI: | http://hdl.handle.net/10261/288847 | DOI: | 10.1016/S0143-4160(98)90079-3 | ISSN: | 0143-4160 |
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