Halothane-induced intracellular calcium release in cholinergic cells |
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Authors: | Renato S Gomez Cristina Guatimosim Jos Barbosa Jr Andr R Massensini Marcus V Gomez Marco A M Prado |
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Institution: | Renato S. Gomez, Cristina Guatimosim, José Barbosa, Jr , André R. Massensini, Marcus V. Gomez,Marco A. M. Prado |
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Abstract: | Low concentrations of halothane and isoflurane can release acetylcholine in an extracellular Ca2+-independent manner. In the present study, a cholinergic cell line (SN56) was used to examine whether release of calcium from intracellular stores occurs in the presence of halothane. Changes in intracellular calcium concentration (Ca2+]i) were measured using fluo-3, a fluorescent calcium-sensitive dye and laser scanning confocal microscopy. Halothane, at sub-anesthetic concentrations (14, 28, 40 and 56 μM), increased Ca2+]i in SN56 cells. This effect remained even when the cells were perfused with medium lacking extracellular calcium, suggesting the involvement of intracellular Ca2+ sources. SN56 cells responded to ryanodine by increasing Ca2+]i and this effect was blocked by dantrolene, an inhibitor of Ca2+-release from ryanodine-sensitive stores. The effect of halothane was attenuated after the increase in Ca2+]i induced by ryanodine and it was suppressed by dantrolene, suggesting the participation of ryanodine-sensitive stores. Using cyclopiazonic acid, a Ca2+-ATPase inhibitor, we investigated whether the depletion of intracellular Ca2+ stores interfered with the effect of halothane. Cyclopiazonic acid significantly decreased the increase in Ca2+]i induced by the volatile anesthetic. It is suggested that sub-anesthetic concentrations of halothane may increase Ca2+]i by releasing Ca2+ from intracellular stores in cholinergic cells. |
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Keywords: | SN56 cell Halothane Intracellular calcium Fluo-3 Ryanodine |
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