title: Control of acid secretion in cultured gas gland cells of the European eel Anguilla anguilla.
abstract: Single cells and cell clusters isolated from the swimbladder epithelium of the European eel Anguilla anguilla attached to collagen S-coated petri dishes and proliferated in a modified Dulbecco's modified Eagle's medium, supplemented with 0.5% fetal calf serum. At a temperature of 20-22 degrees C, the growing colonies reached confluence typically within 6-8 days. Activities of glycolytic and pentose phosphate shunt enzymes remained stable or increased only slightly during the first 10 days of primary culture. Incubated in a defined medium providing glucose as a fuel, gas gland cells in primary culture produced and released lactic acid. The rate of acid secretion of cultured gas gland cells measured with a cytosensor microphysiometer was not influenced by cholinergic stimulation. Similarly, the Ca2+ ionophore A-23187 had no effect. Adrenergic stimulation with epinephrine or the beta-agonist isoproterenol also did not increase the rate of acid secretion, indicating that in gas gland cells the metabolic activity cannot be stimulated via beta-adrenergic stimulation followed by an increase in adenosine 3',5'-cyclic monophosphate (cAMP). Artificially increasing the intracellular concentration of cAMP by incubation with forskolin or the cAMP analogue 8-(4-chlorophenylthio)-cAMP even resulted in a marked reduction in the rate of acid secretion. The results demonstrate that primary cell culture provides a useful means for the analysis of metabolic control and of ion transfer processes in swimbladder gas gland cells.
authors.affiliation: Institut für Physiologie, Ruhr-Universität Bochum, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Pelster
authors.forename: L
authors.initials: L
authors.lastname: Pott
mesh_terms: D000143:Acids / Q000378:metabolism*; D000322:Adrenergic Agonists / Q000494:pharmacology; D000400:Air Sacs / Q000378:metabolism*; D000811:Anguilla / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D004837:Epinephrine / Q000494:pharmacology; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism; D007545:Isoproterenol / Q000494:pharmacology
chemical_list: D000143:Acids; D000322:Adrenergic Agonists; D007545:Isoproterenol; D004837:Epinephrine