Contribution of electrogenic sodium-potassium ATPase to resting membrane potential of cultured rat skeletal myotubes.

The contribution of electrogenic Na+ -K+ ATPase to resting membrane potential (Em) of mature and developing rat skeletal myotubes in culture was determined by examining effects of inhibition of this enzyme on Em. Ouabain, a specific Na+-K+ ATPase inhibitor, caused resting Em to decrease within 30 s by 5-8 mV and reach a minimum value of about -60 mV after 5 min. The decrease in Em was not accompanied by a decrease in input resistance for up to 15 min after application. Resting Em was found to be dependent on the temperature of the recording medium with maximum values of Em ranging from -85 to -90 mV at a temperature of 35-37 degrees C and minimum values about -60 mV at 10-15 degrees C. Ouabain (1 mM), added to cultures at low temperature (10-15 degrees C) did not further decrease Em but did prevent the increase in Em that occurs with increasing temperature up to 37 degrees C. Resting Em of cultured myotubes was reduced to about -60 mV by reducing the supply of ATP either with 2,4 dinitrophenol (DNP), which inhibits oxidative phosphorylation or with fluorodinitrobenzene (FDNB), which inhibits creatine phosphokinase. Neither of these compounds, when added to cultures in the presence of ouabain, reduced resting Em to a value lower than that obtained with ouabain alone.(ABSTRACT TRUNCATED AT 250 WORDS)