Fluoroaluminate activation of different components of the calcium signal in an exocrine cell.

In isolated cells from the avian supra-orbital nasal gland, used as a model for exocrine ion secretion, addition of NaF (2-15 mM) produced a slow Al3(+)-enhanced increase in intracellular Ca2+ concn. ([Ca2+]i), resulting in a more than 2-fold sustained elevation in [Ca2+]i. Simultaneously, cellular Ins(1,4,5)P3 contents became markedly elevated, suggesting an AlF4- activation of a phospholipase C-specific G-protein. Subsequent addition of the muscarinic agonist carbachol failed to produce any further sustained increase in [Ca2+]i, indicating that the AlF4(-)-induced increase in [Ca2+]i involves a Ca2(+)-entry pathway identical with that activated by carbachol. In low-Ca2+ media (extracellular [Ca2+] = 0.04 mM) no such increase in [Ca2+]i, either sustained or transient, is seen, although cellular Ins(1,4,5)P3 levels were markedly elevated. Despite the failure to observe any change in [Ca2+]i in the low-Ca2+ medium, estimation of the size of the agonist-sensitive Ca2+ stores (determined as the magnitude of the transient change in [Ca2+]i induced by carbachol) revealed that these are progressively emptied by the action of AlF4-. However, the onset of this emptying showed an initial lag period of at least 2 min (with 5 mM-NaF plus 10 microM-AlCl3). In marked contrast, determinations of the magnitude of the Ca2(+)-entry pathway under identical conditions showed that this was significantly activated after as little as 1 min of AlF4- treatment. This suggests that, under these conditions, activation of Ca2+ entry in these cells preceded the release of Ca2+ from agonist-sensitive stores, contradicting current models in which the receptor-enhanced entry of extracellular Ca2+ is entirely dependent on, and subsequent to, the prior release of Ca2+ from the intracellular stores.