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Department of Physiology, University of Massachusetts Medical School, Worcester, Massachusetts 01655
In airway epithelial cells, extracellular ATP (ATPo) stimulates an initial transient increase in intracellular Ca2+ concentration that is followed by periodic increases in intracellular Ca2+ concentration (Ca2+ oscillations). The characteristics and mechanism of these ATP-induced Ca2+ responses were studied in primary cultures of rabbit tracheal cells with digital video fluorescence microscopy and the Ca2+-indicator dye fura 2. The continual presence of ATPo at concentrations of 0.1-100 µM stimulated Ca2+ oscillations that persisted for 20 min. The frequency of the Ca2+ oscillations was found to be dependent on both ATPo concentration and intrinsic sensitivity of each cell to ATPo. Cells exhibited similar Ca2+ oscillations to extracellular UTP (UTPo), but the oscillations typically occurred at lower UTPo concentrations. The ATP-induced Ca2+ oscillations were abolished by the phospholipase C inhibitor U-73122 and by the endoplasmic reticulum Ca2+-pump inhibitor thapsigargin but were maintained in Ca2+-free medium. These results are consistent with the hypothesis that in airway epithelial cells ATPo and UTPo act via P2U purinoceptors to stimulate Ca2+ oscillations by the continuous production of inositol 1,4,5-trisphosphate and the oscillatory release of Ca2+ from internal stores. ATP-induced Ca2+ oscillations of adjacent individual cells occurred independently of each other. By contrast, a mechanically induced intercellular Ca2+ wave propagated through a field of Ca2+-oscillating cells. Thus Ca2+ oscillations and propagating Ca2+ waves are two fundamental modes of Ca2+ signaling that exist and operate simultaneously in airway epithelial cells.
inositol 1,4,5-trisphosphate; calcium waves; uridine 5'-triphosphate; adenosine 5'-triphosphate; cell signaling
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