RT Journal Article SR Electronic T1 Cholera Toxin-Stimulated Cyclic AMP Accumulation in Glial Tumor Cells JF Molecular Pharmacology JO Mol Pharmacol FD American Society for Pharmacology and Experimental Therapeutics SP 59 OP 67 VO 20 IS 1 A1 MARGARET A. BROSTROM A1 CHARLES O. BROSTROM A1 SU-CHEN HUANG A1 DONALD J. WOLFF YR 1981 UL http://molpharm.aspetjournals.org/content/20/1/59.abstract AB Accumulation of cyclic AMP in response to cholera toxin was studied in Ca2+-depleted and Ca2+-restored C6 glial tumor cells. Accumulation was retarded when the cells were depleted of Ca2+, but the maximal cyclic AMP content observed with toxin in Ca2+-restored cells was eventually attained in Ca2+-depleted cells. The concentration of cholera toxin which elicited half-maximal cyclic AMP accumulation (apparent Kact) was not altered by Ca2+. Micromolar free Ca2+ concentrations in the extracellular medium were sufficient to overcome the retardation of toxin-stimulated cyclic AMP accumulation in Ca2+-depleted cells. The effect of Ca2+ on cyclic AMP accumulation in response to toxin was rapid and was reversible upon the addition of ethylene glycol bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) in excess of the cation. Accumulation of cyclic AMP by intact cells correlated with increases in the adenylate cyclase activities of Ca2+-depleted and Ca2+-restored cells exposed to cholera toxin. Ca2+, therefore, appears to enhance the rate of adenylate cyclase activation by cholera toxin. Trifluoperazine retarded cholera toxin-dependent cyclic AMP accumulation in a manner similar to cellular Ca2+ depletion. Cyclic AMP accumulation in both Ca2+-depleted and Ca2+-restored preparations was slowed by the drug at micromolar concentrations. Once maximal cell cyclic AMP was attained, however, neither trifluoperazine nor EGTA in excess of Ca2+ affected the nucleotide contents of cells or their extracellular fluids. Binding of 125I-labeled cholera toxin by intact cells was not altered by Ca2+ or by trifluoperazine. The Ca2+ dependence of cyclic AMP accumulation in response to norepinephrine was not affected by pretreatment of cells with cholera toxin, although the extent of the response to the catecholamine was reduced. Similar effects of Ca2+ on cyclic AMP accumulation in cholera toxin-treated cells were identified in neuroblastoma, neuroblastoma-glioma, pituitary tumor, and Chinese hamster ovary cells.