Multiple calcium pathways induce the expression of SNAP-25 protein in chromaffin cells

Incubation of bovine adrenal chromaffin cells in high K+ (38 mM) during 24- 48 h enhanced 2.5 to five times the expression of SNAP-25 protein and mRNA, respectively. This increase was reduced 86% by furnidipine 9(an L-type Ca2 + channel blocker) but was unaffected by either w-conotoxin GVIA (an N-type Ca2+ channel blocker) or w-agatoxin IVA (a P/Q-type Ca2+ channel blocker). Combined blockade of N and P/Q channels with w-conotoxin MVIIC did, howeve r, block by 76% the protein expression. The inhibitory effects of furnidipi ne were partially reversed when the external Ca2+ concentration was raised from 1.6 to 5 mM. These findings, together with the fact that nicotinic rec eptor activation or Ca2+ release from internal stores also enhanced SNAP-25 protein expression, suggest that an increment of cytosolic Ca2+ concentrat ion ([Ca2+],), rather than its source or Ca2+ entry pathway, is the critica l signal to induce the protein expression, The greater coupling between L-t ype Ca2+ channels and protein expression might be due to two facts: (a) L c hannels contributed 50% to the global [Ca2+], rise induced by 38 mM K+ in i ndo-1-loaded chromaffin cells and (b) L channels undergo less inactivation than N or P/Q channels on sustained stimulation of these cells.