Measurement of both Cyclic [3H]AMP and Cyclic [3H]GMP in Cultured Vascular Smooth Muscle Cells Labeled with [3H]Hypoxanthine: Use in Studies of Cardiovascular Drugs

We describe a method for prelabeling cultured vascular smooth muscle cells that permits rapid and accurate measurements of changes in the amounts of cyclic AMP and of cyclic GMP in 5 x 10(5) cells. This procedure utilizes [3H]hypoxanthine to radiolabel both the adenine and guanine nucleotide pools and simple column chromatographic steps to isolate and separate the 3H-labeled cyclic nucleotides. The application of the method to studies of the …