Biosynthesis of proteins of large dense-core vesicles in rat PC12 cells: regulation by forskolin and phorbol ester.

We have investigated the influence of various second messengers on the biosynthesis of large dense-core vesicle constituents in rat PC12 cells. After treatment with forskolin, phorbol ester or a combination of both substances for up to six days, the messenger RNA levels of several vesicle components were determined by northern blotting. Forskolin increased the expression of messenger RNA encoding the soluble proteins chromogranin B, neuropeptide Y and VGF. Addition of phorbol ester markedly enhanced the effects of forskolin. On the other hand, the expression of two further soluble proteins, chromogranin A and secretogranin II, remained fairly unchanged with all treatments tested. Amongst partly membrane-bound vesicle components, the biosynthesis of glycoprotein III and peptidylglycine alpha-amidating mono-oxygenase was significantly up-regulated by combined treatment with forskolin plus phorbol ester. The carboxypeptidase H messenger RNA increased due to phorbol ester and after long-term application of both drugs. In contrast, phorbol ester alone or plus forskolin down-regulated the expression of dopamine beta-hydroxylase. Essentially the same applies to the intrinsic membrane protein cytochrome b-561, whose messenger RNA level declined in all treatment groups. In conclusion, our results show that forskolin and phorbol ester can regulate the composition of large dense-core vesicles in quite distinct patterns.