Pertussis toxin stimulates the secretion of [Met5]-enkephalin and the expression of proenkephalin A mRNA in bovine adrenal medullary chromaffin cells.
[Met5]-Enkephalin (ME) secretion and the expression of proenkephalin A (proENK) mRNA were studied following long-term exposure of bovine adrenal medullary chromaffin (BAMC) cells to pertussis toxin. Treatment with pertussis toxin for 24 h increased the secretion of ME in a concentration- and time-dependent manner. The magnitude of ME secretion continued to increase with time in the presence of pertussis toxin. The intracellular concentration of ME in the pertussis toxin-treated group was not significantly different from controls, suggesting that elevated levels of ME secretion result from increased biosynthesis of ME rather than from release of stored ME. Prolonged (24 h) stimulation of BAMC cells with pertussis toxin also increased proENK gene expression. Pretreatment with nimodipine (a calcium channel blocker) and calmidazolium (a calmodulin antagonist) inhibited both the secretion of ME and the increase in proENK mRNA levels induced by pertussis toxin, while the intracellular calcium antagonist dantrolene and the protein kinase C inhibitors sphingosine and H7 [1-(5-isoquinolinylsulfonyl)-2-methylpiperazine] were ineffective in blocking pertussis toxin-induced responses. Forskolin (an adenyl cyclase activator) and isobutyl methyl xanthine (a phosphodiesterase inhibitor) increased both ME secretion and proENK mRNA levels; pertussis toxin synergistically increased the secretion of ME with these cyclic AMP-elevating agents but had only an additive effect with these agents on the level of proENK mRNA. Our results suggest that a pertussis toxin-sensitive G protein may tonically regulate the secretion of ME as well as the level of proENK mRNA.(ABSTRACT TRUNCATED AT 250 WORDS)
Suh
H H
HH
Neuropharmacology Section, National Institute of Environmental Health Science, National Institute of Health, Research Triangle Park, N.C. 27709.
Hudson
P
P
McMillian
M K
MK
Hong
J S
JS
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D037361:Adenylate Cyclase Toxin; D000313:Adrenal Medulla / Q000187:drug effects* / Q000502:physiology; D000818:Animals; D001483:Base Sequence; D002147:Calmodulin / Q000037:antagonists & inhibitors; D002417:Cattle; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D015342:DNA Probes; D003620:Dantrolene / Q000494:pharmacology; D004744:Enkephalin, Methionine / Q000378:metabolism*; D004745:Enkephalins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D007093:Imidazoles / Q000494:pharmacology; D008969:Molecular Sequence Data; D009553:Nimodipine / Q000494:pharmacology; D037342:Pertussis Toxin; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015290:Second Messenger Systems / Q000187:drug effects / Q000502:physiology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology*
D037361:Adenylate Cyclase Toxin; D002147:Calmodulin; D015342:DNA Probes; D004745:Enkephalins; D007093:Imidazoles; D011498:Protein Precursors; D012333:RNA, Messenger; D010566:Virulence Factors, Bordetella; C029992:proenkephalin; D005576:Colforsin; C031938:calmidazolium; D009553:Nimodipine; D004744:Enkephalin, Methionine; D037342:Pertussis Toxin; D003620:Dantrolene; D015056:1-Methyl-3-isobutylxanthine