Flash photolysis studies of the localization of calcium release sites in rat parotid isolated acinar cells.

1. The temporal relationship between cytosolic free Ca2+ concentration ([Ca2+]i) and activation of membrane current responses in single rat parotid acinar cells has been examined. Activation of muscarinic receptors by carbachol (CCh) at -40 mV (midway between EK and ECl under our experimental conditions) frequently evoked biphasic current responses, application of 2 microM CCh leading to rapid activation of an inward current followed by a slower outward current. 2. Photochemical release of inositol 1,4,5-trisphosphate (InsP3), from 'caged' InsP3, by a brief near-UV flash, evoked similar biphasic current responses at -40 mV. In contrast, elevation of [Ca2+]i by photolysis of the caged calcium compound nitr-5 at -40 mV activated only monophasic current responses. 3. These results can be explained by a model in which the InsP3-sensitive Ca2+ release sites are localized at the luminal pole of the cell, combined with a relative preponderance of Ca(2+)-activated Cl- channels at that pole, and a relative preponderance of Ca(2+)-activated K+ channels at the basal end.