Time Course of Release of Catecholamine and Other Granular Contents from Perifused Adrenal Chromaffin Cells of Guinea-pig

Overview

Journal J Physiol

Specialty Physiology

Date 1983 Aug 1

PMID 6620178

Citations 4

Authors

S Ito

Affiliations

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Abstract

Experiments were carried out to investigate the time course of the release of catecholamine, dopamine-beta-hydroxylase (DBH) and adenine nucleotides from isolated chromaffin cells of guinea-pig adrenal gland. When the isolated chromaffin cells were incubated with medium containing acetylcholine (ACh) (0.1 mM), veratridine (0.1 mM) or scorpion (Leiurus quinquestriatus) venom, (10 micrograms/ml.), catecholamine was released into the medium. Catecholamine secretion induced by veratridine or scorpion venom was inhibited by tetrodotoxin (1 microM) but not by atropine (0.1 mM) plus hexamethonium (0.1 mM). On the other hand, the secretory response to ACh was abolished by the cholinergic blocking drugs but not by tetrodotoxin. DBH was released together with catecholamine into the medium in which cells were suspended with these drugs. The ratio of catecholamine (n-mole) to DBH activity (n-mole/hr) appearing in the supernatant was 7.08 +/- 0.55, 6.60 +/- 0.27 and 8.91 +/- 0.47 for ACh, veratridine and scorpion venom, respectively. These values were close to that found in the lysate of chromaffin granules obtained from guinea-pig adrenal glands (7.37 +/- 0.39). The application of ACh or veratridine to perifused chromaffin cells was found to cause a parallel increase in catecholamine and DBH secretion in the perifusion medium without corresponding amounts of phenylethanolamine-N-methyltransferase leakage. However, DBH secretion tended to last for a longer period than catecholamine secretion. Adenine nucleotides were released from perifused chromaffin cells together with catecholamine, by ACh and veratridine. ATP added to the perifusion medium was metabolized to ADP and AMP, of which the ratio (ATP, 21.6%; ADP, 34%; AMP, 17.9%) was close to those of adenine nucleotides released from the cells. The secretion of adenine nucleotides induced by both secretagogues ceased much faster than the catecholamine secretion, so that molar ratio of catecholamine to adenine nucleotides was gradually increased during and after stimulation. The results indicate that catecholamine secretion is accompanied with a simultaneous release of DBH and ATP from adrenal chromaffin cells. Therefore, it is suggested that the delayed output of DBH, unlike catecholamine secretion, in perfused adrenal glands results from the presence of a diffusion barrier for this protein. The releasable secretory granules of isolated chromaffin cells are suggested to be heterogeneous with respect to the ratio of catecholamine to ATP.

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References

DOUGLAS W, Poisner A, Rubin R . Efflux of adenine nucleotides from perfused adrenal glands exposed to nicotine and other chromaffin cell stimulants. J Physiol. 1965; 179(1):130-7. PMC: 1357327. DOI: 10.1113/jphysiol.1965.sp007652. View

DOUGLAS W, Poisner A . Evidence that the secreting adrenal chromaffin cell releases catecholamines directly from ATP-rich granules. J Physiol. 1966; 183(1):236-48. PMC: 1357539. DOI: 10.1113/jphysiol.1966.sp007863. View

DOUGLAS W, Poisner A . On the relation between ATP splitting and secretion in the adrenal chromaffin cell: extrusion of ATP (unhydrolysed) during release of catecholamines. J Physiol. 1966; 183(1):249-56. PMC: 1357540. DOI: 10.1113/jphysiol.1966.sp007864. View

Banks P . The release of adenosine triphosphate catabolites during the secretion of catecholamines by bovine adrenal medulla. Biochem J. 1966; 101(2):536-41. PMC: 1270137. DOI: 10.1042/bj1010536. View

Smith A, Winkler H . A simple method for the isolation of adrenal chromaffin granules on a large scale. Biochem J. 1967; 103(2):480-2. PMC: 1270431. DOI: 10.1042/bj1030480. View

Kirshner N, Sage H, Smith W . Mechanism of secretion from the adrenal medulla. II. Release of catecholamines and storage vesicle protein in response to chemical stimulation. Mol Pharmacol. 1967; 3(3):254-65. View

BLASCHKO H, COMLINE R, SCHNEIDER F, Silver M, Smith A . Secretion of a chromaffin granule protein, chromogranin, from the adrenal gland after splanchnic stimulation. Nature. 1967; 215(5096):58-9. DOI: 10.1038/215058a0. View

LADURON P, Belpaire F . Tissue fractionation and catecholamines. II. Intracellular distribution patterns of tyrosine hydroxylase, dopa decarboxylase, dopamine-beta-hydroxylase, phenylethanolamine N-methyltransferase and monoamine oxidase in adrenal medulla. Biochem Pharmacol. 1968; 17(7):1127-40. DOI: 10.1016/0006-2952(68)90048-8. View

Molinoff P, Weinshilboum R, Axelrod J . A sensitive enzymatic assay for dopamine- -hydroxylase. J Pharmacol Exp Ther. 1971; 178(3):425-31. View

10.

Joh T, Goldstein M . Isolation and characterization of multiple forms of phenylethanolamine N-methyltransferase. Mol Pharmacol. 1973; 9(1):117-29. View

11.

Aberg H, Hansson H, Wetterberg L, Ross S, Froden O . Dopamine-beta-hydroxylase in human lymph. Life Sci. 1974; 14(1):65-71. DOI: 10.1016/0024-3205(74)90246-x. View

12.

Lastowecka A, TRIFARO J . The effect of sodium and calcium ions on the release of catecholamines from the adrenal medulla: sodium deprivation induces release by exocytosis in the absence of extracellular calcium. J Physiol. 1974; 236(3):681-705. PMC: 1350857. DOI: 10.1113/jphysiol.1974.sp010460. View

13.

Cubeddu L, Barnes E, Langer S, WEINER N . Release of norepinephrine and dopamine-beta-hydroxylase by nerve stimulation. I. Role of neuronal and extraneuronal uptake and of alpha presynaptic receptors. J Pharmacol Exp Ther. 1974; 190(3):431-50. View

14.

DIXON W, Garcia A, KIRPEKAR S . Release of catecholamines and dopamine beta-hydroxylase from the perfused adrenal gland of the cat. J Physiol. 1975; 244(3):805-24. PMC: 1330837. DOI: 10.1113/jphysiol.1975.sp010827. View

15.

Lagercrantz H, Fried G, Dahlin I . An attempt to estimate the in vivo concentrations of noradrenaline and ATP in sympathetic large dense core nerve vesicles. Acta Physiol Scand. 1975; 94(1):136-8. DOI: 10.1111/j.1748-1716.1975.tb05871.x. View

16.

Hochman J, Perlman R . Catecholamine secretion by isolated adrenal cells. Biochim Biophys Acta. 1976; 421(1):168-75. DOI: 10.1016/0304-4165(76)90180-x. View

17.

Peer L, Winkler H, Snider S, GIBB J, Baumgartner H . Synthesis of nucleotides in adrenal medulla and their uptake into chromaffin granules. Biochem Pharmacol. 1976; 25(3):311-5. DOI: 10.1016/0006-2952(76)90220-3. View

18.

VAN ORDEN L, Burke J, Redick J, Rybarczyk K, Van Orden D, Baker H . Immunocytochemical evidence for particulate localization of phenylethanolamine-N-methyltransferase in adrenal medulla. Neuropharmacology. 1977; 16(2):129-33. DOI: 10.1016/0028-3908(77)90060-0. View

19.

UVNAS B, Aborg C . The ability of ATP-free granule material from bovine adrenal medulla to bind inorganic cations and biogenic amines. Acta Physiol Scand. 1977; 99(4):476-83. DOI: 10.1111/j.1748-1716.1977.tb10401.x. View

20.

Van Dyke K, Robinson R, Urquilla P, Smith D, Taylor M, Trush M . An analysis of nucleotides and catecholamines in bovine medullary granules by anion exchange high pressure liquid chromatography and fluorescence. Evidence that most of the catecholamines in chromaffin granules are stored without associated ATP. Pharmacology. 1977; 15(5):377-91. DOI: 10.1159/000136714. View