Differential accumulation of catecholamines, proenkephalin- and chromogranin A-derived peptides in the medium after chronic nicotine stimulation of cultured bovine adrenal chromaffin cells.
We have used an antiserum to a synthetic peptide fragment of bovine chromogranin A (ChrgA)[Tyr0] bovine ChrgA (306-313): YLSKEWEDA, together with antibodies to proenkephalin-derived peptides, to measure the release of immunoreactive peptides from nicotine-stimulated cultured bovine adrenal chromaffin cells. Over a period of 6 hr the accumulation of YLSKEWEDA immunoreactivity and Met-enkephalin Arg6Gly7Leu8 (MERGL) immunoreactivity in the medium of 10 microM nicotine-stimulated cells was shown to be biphasic; the initial phase occurred in the first 15-30 min and the second phase reached a peak after 4 hr. In contrast, catecholamine release occurred monophasically over the initial 15-30 min. Investigation of the second phase of peptide accumulation revealed that it was due in part to nicotine-evoked exocytosis and in part to extracellular processing of high molecular weight precursor proteins.
Watkinson
A
A
MRC Secretory Control Research Group, University of Liverpool, U.K.
O'Sullivan
A J
AJ
Burgoyne
R D
RD
Dockray
G J
GJ
D000313:Adrenal Medulla / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D000818:Animals; D002395:Catecholamines / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D002850:Chromatography, Gel; D053379:Chromogranin A; D002864:Chromogranins / Q000378:metabolism*; D003470:Culture Media; D004745:Enkephalins / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009538:Nicotine / Q000494:pharmacology*; D010446:Peptide Fragments / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism*; D011863:Radioimmunoassay; D013997:Time Factors
D002395:Catecholamines; D053379:Chromogranin A; D002864:Chromogranins; D003470:Culture Media; D004745:Enkephalins; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; D011498:Protein Precursors; C029992:proenkephalin; D009538:Nicotine