Abstract
A method has been devised for measuring the level of adenosine 3', 5'-monophosphate (cyclic AMP) newly synthesized ruring in vitro incubation of the isolated rat kidney cells with adenine-8-14C. By combined use of the BaSO4 treatment and the onedimensional ascending cellulose thin-layer chromatography with stepwise development by two solvents, newly formed or labeled cyclic AMP (14C-cyclic AMP) could completely be separated from the related radioactive metabolites in the cells.Effect of lysine-vasopressin on the isolated rat kidney cells was studied by using the radioassay method. It was shown that (1) in the presence of theophylline, vasopressin increased 14C-cyclic AMP level, and (2) the stimulative effect was specific to vasopressin. Moreover, (3) vasopressin was shown to increase 14C-cyclic AMP level markedly in the medium but little in the cells. It was thus indicated that over-all synthesis of cyclic AMP in the rat kidney cells and consequential excretion of the newly synthesized cyclic AMP into the medium were enhanced by vasopressin.
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