Effect of protein synthesis inhibitors on synexin levels and secretory response in bovine adrenal medullary chromaffin cells.
The effects of the protein synthesis inhibitors actinomycin D and cycloheximide on the cellular content of the calcium binding protein synexin, and on the secretory response of cultured bovine adrenal medullary chromaffin cells were determined. Both protein synthesis inhibitors produced a slow decrease in the cellular synexin content. The synexin level was reduced by 50% after 133 h of incubation in the presence of 2 micrograms/ml actinomycin D or 5 micrograms/ml cycloheximide. However, this was partly due to an artefactual stabilization of synexin, since metabolic labelling of synexin with [35S]methionine showed that the half-time of degradation was only 40 h. The secretory response of chromaffin cells was quickly diminished in the presence of protein synthesis inhibitors. Catecholamine secretion induced by membrane depolarization or barium stimulation of intact cells, or by calcium stimulation of digitonin-permeabilized cells was decreased by 77-82% after 24 h of incubation in the presence of 5 micrograms/ml cycloheximide. These results suggest that, in addition to synexin, at least one or more proteins with a shorter half-time of degradation than synexin are involved in the secretory response of adrenal chromaffin cells.
Cardenas
A M
AM
Laboratory of Cell Biology and Genetics, NIDDK, NIH, Bethesda, MD 20892-0840.
Kuijpers
G A
GA
Pollard
H B
HB
D000313:Adrenal Medulla / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D000704:Analysis of Variance; D000818:Animals; D017310:Annexin A7 / Q000096:biosynthesis / Q000378:metabolism*; D017609:Barium Compounds / Q000494:pharmacology; D001483:Base Sequence; D002122:Calcium Chloride / Q000494:pharmacology; D002395:Catecholamines / Q000378:metabolism*; D002417:Cattle; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002712:Chlorides / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology*; D003609:Dactinomycin / Q000494:pharmacology*; D007031:Hypothalamus; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D008969:Molecular Sequence Data; D009474:Neurons; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D016716:PC12 Cells; D011189:Potassium Chloride / Q000494:pharmacology; D051381:Rats; D014529:Uridine / Q000378:metabolism
D017310:Annexin A7; D017609:Barium Compounds; D002395:Catecholamines; D002712:Chlorides; D016376:Oligonucleotides, Antisense; C024986:barium chloride; D003609:Dactinomycin; D011189:Potassium Chloride; D003513:Cycloheximide; D007930:Leucine; D002122:Calcium Chloride; D014529:Uridine