Measurement of the change in cell membrane capacitance (Cm) along with the change in IK(Ca) was used to investigate the effects of bradykinin and caffeine on the secretory process in rat adrenal chromaffin cells. In a Ca2+-free external solution, bradykinin (100 nM) caused a transient increase in Cm with a concurrent change in IK(Ca). Extracellular application of neomycin as an inhibitor of phospholipase C activity reversibly inhibited the bradykinin-activated event, implying an IP3-mediated increase of submembrane-free Ca2+. The increases in Cm and IK(Ca) caused by bradykinin were transient even with the sustained application of bradykinin. Caffeine also caused exocytosis in the Ca2+-free solution, and this was irreversibly blocked by ryanodine (1 microM) in a use-dependent manner. Caffeine-sensitive intracellular Ca2+ stores were also depleted in several seconds and recovered by an influx of external Ca2+. The sequential application of bradykinin and caffeine showed that these are likely to activate Ca2+ release from the same or distinct but rapidly equilibrating intracellular Ca2+ stores. The single cell assay of exocytosis and the increase in IK(Ca) revealed cell-to-cell variability in bradykinin- and caffeine-induced exocytotic response. Our results suggest that Ca2+ release from intracellular stores potentially increases submembrane Ca2+ concentration and modulates simultaneously two submembrane Ca2+-dependent processes, exocytosis and IK(Ca), in rat adrenal chromaffin cells.
Copyright 1998 Elsevier Science B.V. All rights reserved.