TEMPORAL RELATIONSHIPS BETWEEN CA2+ STORE MOBILIZATION AND CA2+ ENTRY IN AN EXOCRINE CELL

Consideration of the principal current models for agonist-induced activation of Ca2+ entry in electrically non-excitable cells suggests that it may be possible to distinguish between them on the basis of predicted differences in the temporal relationship(s) between intracellular Ca2+ release and the activation of Ca2+ entry. Measurements of changes in [Ca2+](i) and Mn2+ quench in individual exocrine cells from the avian nasal grand indicate that, whereas Ins(1,4,5)P-3-induced release of intracellular Ca2+ occurs within 3-5 s, the increase in Mn2+ quench is delayed by some 20-30 s. Mn2+ quench rate is similarly increased by thapsigargin, and is blocked by SK&F 96365, indicating that the increased Mn2+ quench observed genuinely reflects agonist-enhanced activity of the divalent cation entry pathway normally traversed by Ca2+ Additional experiments indicate that the observed delay is not due to inhibition of this pathway by elevated [Ca2+](i). Furthermore, the delay cannot be explained by the time required for Ins(1,3,4,5)P-4 generation, which is essentially maximal within 10 s of agonist addition. It is concluded that the observed delay in the activation of the Ca2+ entry pathway is best explained by 'capacitative' models where increased entry requires the generation, and transmission to the plasma membrane, of an unknown messenger as a direct result of the depletion of intracellular Ca2+ stores.