Differential accumulation of catecholamines, proenkephalin- and chromogranin A-derived peptides in the medium after chronic nicotine stimulation of cultured bovine adrenal chromaffin cells

• Published in: Peptides (New York, N.Y. : 1980) Vol. 11; no. 3; pp. 435 - 441
• Main Authors: Watkinson, A., O'Sullivan, A.J., Burgoyne, R.D., Dockray, G.J.
• Format: Journal Article
• Language: English
• Published: New York, NY Elsevier Inc 01.05.1990; Elsevier Science
• Subjects: Adrenal Medulla - cytology; Adrenal Medulla - drug effects; Adrenal Medulla - metabolism; Adrenals. Interrenals; Adrenomedullary hormones. Regulation; Animals; Biological and medical sciences; Catecholamines; Catecholamines - metabolism; Cattle; Cells, Cultured; Chromaffin cells; Chromatography, Gel; Chromogranin A; Chromogranins - metabolism; Culture Media; Enkephalins; Enkephalins - metabolism; Fundamental and applied biological sciences. Psychology; Nerve Tissue Proteins - metabolism; Nicotine - pharmacology; Peptide Fragments - metabolism; Postsecretory processing; Protein Precursors - metabolism; Radioimmunoassay; Time Factors; Vertebrates: endocrinology; Chromaffin cells; Catecholamines; Postsecretory processing; Chromogranin A; Enkephalins; Cell culture; Endocrine gland; Adrenal glands; Secretion; Enkephalin; Ox; Catecholamine; Chromaffin cell; In vitro; Vertebrata; Mammalia; Adrenal hormone; Artiodactyla; Ungulata; Nicotine
• ISSN: 0196-9781; 1873-5169
• DOI: 10.1016/0196-9781(90)90039-8

We have used an antiserum to a synthetic peptide fragment of bovine chromogranin A (ChrgA)[Tyr o] 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 Arg 6Gly 7Leu 8 (MERGL) immunoreactivity in the medium of 10 μM 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.