Temperature Sensitivity of Cyclic AMP Production and Catecholamine-Induced Refractoriness in a Rat Astrocytoma Cell Line

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 75; no. 11; pp. 5520 - 5524
• Main Authors: Nickols, G. Allen, Brooker, Gary
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences of the United States of America 01.11.1978; National Acad Sciences
• Subjects: 1-Methyl-3-isobutylxanthine - pharmacology; Agonists; Astrocytoma; Biological Sciences: Cell Biology; Catecholamines; Cell culture techniques; Cell Line; Cell lines; Cells; Cultural groups; Cultured cells; Cyclic AMP - metabolism; Cyclic nucleotides; Isoproterenol - pharmacology; Kinetics; Low temperature; Receptors; Temperature; Theophylline - analogs & derivatives
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.75.11.5520

Intracellular cyclic AMP was increased more than 100-fold when rat C6-2B astrocytoma cells were treated with isoproterenol in the cold (4 degrees C). When the cells were treated with the phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine, and isoproterenol, cyclic AMP levels rose more than 150-fold. The levels achieved compared favorably with the 250-fold increase in cyclic AMP produced by (-)isoproterenol at 37 degrees C. (-)Isoproterenol at 5 nM stimulated half-maximal cyclic AMP production at 4 degrees C and at 37 degrees C and was blocked by (-)propranolol at both temperatures. The concentrations of cyclic AMP attained by these cells after (-)isoproterenol stimulation in the cold may be accounted for, in part, by alterations in the efflux of the nucleotide from the cells since extracellular cyclic AMP, an indicator of cyclic AMP efflux, was found to be dramatically reduced in the cold. The cells, when exposed to (-)isoproterenol for up to 6 hr at low temperature, maintained normal responsiveness to this agent when rechallenged at 4 degrees or 37 degrees C. Thus, they did not display agonist-induced refractoriness during that period of exposure at 4 degrees C, although refractoriness is always seen within 90 min at 37 degrees C. Refractoriness, once established by (-)isoproterenol treatment at 37 degrees C, was not reversed by exposure of the cells to cold. These data suggest that the development of catecholamine refractoriness requires a temperature-sensitive step that lies distal to the hormone-receptor interaction and cyclic AMP generation.