Cyclic adenosine 3':5'-monophosphate and cytosolic calcium exert opposing effects on biosynthesis of tetrodotoxin-sensitive sodium channels in rat muscle cells

We have previously presented evidence that electrical activity and increased cytosolic calcium reduce the density of sarcolemmal tetrodotoxin (TTX)-sensitive sodium channels in cultured rat muscle cells (Sherman, S.J., and W.A. Catterall (1984) Proc. Natl. Acad. Sci. U.S.A. 81: 262-266). We show here that growth of cells in ryanodine has a biphasic effect on sodium channel number. At low concentrations (0.3 to 10 μM) where this drug releases calcium from the sarcoplasmic reticulum into the cytoplasm, sodium channel number is reduced 62%; whereas, at higher concentrations where total cellular calcium is depleted, the density of sodium channels is increased 40% above control. These results provide further evidence for modulation of sodium channel number by cytosolic calcium. Growth of muscle cells in the presence of agents that mimic cyclic AMP (cAMP) action or increase intracellular cAMP levels including 8-bromo-cyclic AMP (8-BrcAMP), cyclic nucleotide phosphodiesterase inhibitors, and forskolin increased sodium channel density up to 125%. This action did not involve changes in spontaneous electrical activity. Dibutyryl cGMP had no effect. Measurement of the turnover rate of sodium channels after block of channel accumulation by tunicamycin (1.5 μg/ml) gave a half-time of 18 hr for exponential decay of TTX-sensitive sodium channels in cultured rat muscle cells after an initial 6-hr lag period. Treatments which modulate sodium channel number through changes in cytosolic calcium or cAMP had no effect on the rate of channel turnover. The increase of sodium channel number after inhibition of electrical activity or treatment with 8-BrcAMP was half- maximal at 17 hr, consistent with an increase in the rate of sodium channel biosynthesis and/or incorporation into the sarcolemma without a change in channel turnover time. We conclude that cytosolic calcium decreases and cAMP increases sodium channel number by modulating the rate of biosynthesis and/or processing of channel components. The biochemical mechanisms of these regulatory effects are considered.