An intracellular pool of N-type voltage-operated calcium channels has
recently been described in both IMR32 human neuroblastoma and PC12 rat
pheochromocytoma cells. These channels were found to be
accumulated in subcellular fractions where the chromogranin
B-containing secretory granules were also enriched. Upon
exocytosis N-type calcium channels were reversibly inserted in the
plasma membrane. We have now extended this study to RINm5F rat
insulinoma cells, and characterized the parallelism between the
'regulated' secretion of serotonin and the recruitment of surface
calcium channels. Exocytosis was stimulated by different means,
such as depolarization with high KCl, high Ba2+ alone or
protein kinase C activation; on the other hand exocytosis was inhibited
with the non-selective calcium channel antagonist Cd2+ or
with noradrenaline. Stimulated release was always accompanied,
with parallel kinetics, by calcium channel recruitment, while
inhibition of secretion blocked calcium channel recruitment too.
During repetitive depolarizations we revealed a potentiation of
[Ca2+]i transients in single Fura-2 loaded
RINm5F cells, that was accompanied by an increase in surface VOCCs,
suggesting a physiological role for the newly recruited
channels.
(Click on the figure for an enlarged view.)