Regulation of tyrosine hydroxylase activity in rat PC12 cells by neuropeptides of the secretin family.
Tyrosine hydroxylase, the rate-limiting enzyme in catecholamine biosynthesis, is subject to regulation by the cAMP as well as the calcium and cGMP second messenger systems. Treatment of intact rat PC12 cells with neuropeptides including secretin and vasoactive intestinal polypeptide (VIP) stimulated tyrosine hydroxylase activity 2 to 3-fold in vitro. Secretin (EC50 = 10 nM) was about 3 orders of magnitude more potent than VIP (EC50 = 3 microM). A combination of several protease inhibitors failed to enhance the potency of either peptide. Other members of the secretin family including glucagon and peptide histidine isoleucine (PHI) stimulated tyrosine hydroxylase activity to a lesser extent. Somatostatin, which is not homologous to secretin, was ineffective. The maximal response of tyrosine hydroxylase activation to 1 microM secretin occurred within 6-15 sec. Secretin, VIP, and forskolin also enhanced tyrosine hydroxylase activity (3,4-dihydroxyphenylalanine production) in intact cells, as determined by high performance liquid chromatography and electrochemical detection. Secretin, VIP, PHI, and glucagon increased the levels of cAMP in PC12 cells more than 10-fold, as determined by radioimmunoassay. We also demonstrated that cAMP is released from the cells into the incubation medium following secretin treatment. Secretin and VIP treatment also enhanced the activity of cAMP-dependent protein kinase in a concentration-dependent fashion, as measured subsequently in vitro. Based on the greater potency of secretin in comparison with VIP, PHI, and glucagon, we suggest that the PC12 cells contain a secretin-preferring receptor that increases cAMP levels and brings about an activation of tyrosine hydroxylase activity through the stimulation of cAMP-dependent protein kinase.
Roskoski
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R
Department of Biochemistry and Molecular Biology, Louisiana State University Medical Center, New Orleans.
White
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Knowlton
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Roskoski
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LM
D000310:Adrenal Gland Neoplasms / Q000201:enzymology; D000818:Animals; D002118:Calcium / Q000502:physiology; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000032:analysis; D004295:Dihydroxyphenylalanine / Q000096:biosynthesis; D004789:Enzyme Activation; D010673:Pheochromocytoma / Q000201:enzymology; D051381:Rats; D012633:Secretin / Q000494:pharmacology*; D013806:Theophylline / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis*; D014660:Vasoactive Intestinal Peptide / Q000494:pharmacology
D012633:Secretin; D005576:Colforsin; D014660:Vasoactive Intestinal Peptide; D004295:Dihydroxyphenylalanine; D013806:Theophylline; D000242:Cyclic AMP; D014446:Tyrosine 3-Monooxygenase; D002118:Calcium