Spatial distribution of intracellular, free Ca2+ in isolated rat parotid acini

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Abstract
The spatial distribution of intracellular, free Ca2+ ([Ca2+]i) in rat parotid acini was measured by imaging fura-2 fluorescence from individual acinar cells by means of a digital imaging microscope. Upon cholinergic stimulation in a Krebs-Ringer bicarbonate buffer at (37° C), [Ca2+]i increased synchronously at both the basolateral and luminal membranes as well as in all cells of the secretory endpiece, reaching peak [Ca2+]i levels 1 s after stimulation. Atropine addition caused a rapid down-regulation of [Ca2+]i, which, however, never reached prestimulatory levels. When acini were stimulated in a medium containing 5 nM Ca2+, the Ca2+ mobilization arising from internal pools caused an increase in [Ca2+]i predominantly near the basolateral area, where the endoplasmic reticulum is located, and standing Ca2+ gradients were observed for up to 10 s. A mathematical model is developed to simulate the time courses of the Ca2+ profiles through the cytoplasm using estimated values of the Ca2+ diffusion coefficients and the cytosolic Ca2+ buffering capacity. It is concluded that under physiological conditions, the Ca2+ release from the endoplasmic reticulum is responsible for the activation of the basolaterally located K+ channels. Furthermore, Ca2+ influx from the interstitium is responsible for much of the rise in [Ca2+]i near the luminal membranes, where the Cl− channels are supposed to be located.