BIOSYNTHESIS OF LARGE DENSE-CORE VESICLES IN PC12 CELLS - EFFECTS OF DEPOLARIZATION AND 2ND MESSENGERS ON THE MESSENGER-RNA LEVELS OF THEIR CONSTITUENTS

mRNA levels of various constituents of large dense-core vesicles were determined in PC12 cells during depolarization and/or in the presence of BayK 8644, forskolin or phorbolester. For the soluble (secretory) proteins of the vesicles the mRNAs of chromogranin A and B, secretogranin II, neuropeptide Y and VGF were analyzed. Depolarization in the presence of BayK induced a strong up-regulation of the messages for chromogranin B, neuropeptide Y and VGF. Addition of forskolin enhanced this response for neuropeptide Y and VGF, phorbolester did the same only for VGF. Partly membrane-bound and membrane-spanning components analyzed were carboxypeptidase H, dopamine beta-hydroxylase and glycoprotein III (clusterin), peptidylglycine alpha-amidating mono-oxygenase and cytochrome b-561, respectively. Changes of mRNAs for these components were in general smaller and delayed. Six days of depolarization caused an up-regulation of glycoprotein III, peptidylglycine alpha-amidating mono-oxygenase and carboxypeptidase H mRNA levels which were not further increased by cyclic AMP and phorbolester. The dopamine beta-hydroxylase message increased after 6 days of depolarization, however, addition of phorbolester reduced this effect. For cytochrome b-561 there was no change after any of the conditions employed. These in vitro results are compared with those obtained for the biosynthesis regulation of large dense-core vesicles under in vivo conditions. It is suggested that in vivo acetylcholine and vasoactive intestinal polypeptide released from splanchnic nerve induce a differential change in the biosynthesis of large dense-core vesicles by acting via calcium and protein kinase A and C.