Effect of Insulin on Intracellular pH in Frog Skeletal Muscle Fibers

The effect of insulin on intracellular pH (pH _i ) and membrane potential was studied in frog semitendinosus muscle fibers, using recessed-tip pH-sensitive glass microelectrodes and conventional 3 M KCl-filled microelectrodes. After a lag period of approximately 20 min, insulin [1 mU/ml, 0.1% bovine serum albumin (BSA)] produced a slow hyperpolarization of 2–5 mV and an alkalinization of 0.05–0.10 pH unit, which were both completed within 1 h and were not reversed by washing in insulin-free solution for 1 h. The effect of insulin on the pHi recovery rate from CO ₂ ^- induced acidification was examined at various membrane voltages. At normal membrane voltage, insulin (400 mU/ml, no BSA) slightly increased the slow pH _i recovery (from 0.01 to 0.04 ΔpH/h). In fibers depolarized in 15 mM K to about -50 mV, insulin nearly tripled the recovery rate (from 0.05 to 0.13 ΔpH/h). This insulin-induced recovery was abolished by 1 mM amiloride, a Na-H exchange inhibitor. The increased pH _i recovery in 15 mM K thus represents an increased Na-H exchange, which may be due to an interaction between insulin and either membrane depolarization, per se, or increased intracellular Ca. In fibers depolarized in 50 mM K to about -25 mV, insulin did not affect recovery (0.28 ΔpH/h). This lack of insulin effect might be due to fiber swelling or to the difference in the time course of elevation of intracellular Ca at -25 and -50 mV. These results are consistent with an alkalinizing effect of insulin in frog muscle mediated by Na-H exchange.