Regulation of the chromaffin granule catecholamine transporter in cultured bovine adrenal medullary cells: stimulus-biosynthesis coupling.
The transsynaptic induction of the monoamine transporter present on the membrane of chromaffin granules was studied in primary cultures of dissociated bovine adrenomedullary cells submitted to a chronic secretory stimulation. The amount of the vesicular monoamine transporter was assayed by binding of the specific ligand [3H]-dihydrotetrabenazine. After several days of incubation in the presence of high potassium, the concentration of [3H]-dihydrotetrabenazine binding sites was increased by a 1.5-2.5 factor. This increase was smaller in the presence of the cholinergic agonist carbachol. The long-term inductions of the vesicular monoamine transporter, of tyrosine hydroxylase, and of acetylcholinesterase were of similar magnitude. Under the same conditions, we found no variation in either the activities of other catecholamine biosynthetic enzymes (dopamine beta-hydroxylase and DOPA decarboxylase), or in metabolic enzymes such as lactate dehydrogenase and cytochrome c oxidase, and a decrease in the cellular content of chromogranin A and cytochrome b-561. The induction of the vesicular monoamine transporter was inhibited by the calcium channel antagonists, fluspirilene and nifedipine, and was increased by the agonist Bay K 8644. It was abolished by cycloheximide and actinomycin D. These results indicate that calcium entry into chromaffin cells increases the synthesis of the vesicular monoamine transporter, presumably by transcriptional activation. Elevation of intracellular cyclic AMP concentration or activation of protein kinase C also induced an increase in the expression of the vesicular monoamine transporter. Our results confirm that components of storage vesicle membranes are differentially regulated in response to secretory stimulation, as are several cytosolic or intravesicular soluble proteins.(ABSTRACT TRUNCATED AT 250 WORDS)
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000110:Acetylcholinesterase / Q000032:analysis / Q000378:metabolism; D000313:Adrenal Medulla / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D002217:Carbachol / Q000494:pharmacology; D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism*; D050482:Catecholamine Plasma Membrane Transport Proteins; D002395:Catecholamines / Q000493:pharmacokinetics; D002417:Cattle; D002478:Cells, Cultured; D002837:Chromaffin Granules / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D053379:Chromogranin A; D002864:Chromogranins / Q000032:analysis / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004299:Dopamine beta-Hydroxylase / Q000032:analysis / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism; D026901:Membrane Transport Proteins; D009543:Nifedipine / Q000494:pharmacology; D011188:Potassium / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D013747:Tetrabenazine / Q000031:analogs & derivatives / Q000378:metabolism; D013997:Time Factors; D014316:Tritium; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis / Q000378:metabolism
D002352:Carrier Proteins; D050482:Catecholamine Plasma Membrane Transport Proteins; D002395:Catecholamines; D053379:Chromogranin A; D002864:Chromogranins; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D014316:Tritium; D003609:Dactinomycin; D005576:Colforsin; C032811:dihydrotetrabenazine; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D002217:Carbachol; D003513:Cycloheximide; D000242:Cyclic AMP; D014446:Tyrosine 3-Monooxygenase; D004299:Dopamine beta-Hydroxylase; D011493:Protein Kinase C; D000110:Acetylcholinesterase; D009543:Nifedipine; D011188:Potassium; D013747:Tetrabenazine