Experiments were performed on isolated rat soleus (slow-twitch) and extensor digitorum longus (EDL) (fast-twitch) muscle of 4-week-old rats. In soleus muscle, electrical simulation at 2 Hz for 5 min increased the ouabain-suppressible 86Rb+ uptake by 138%, without significant changes in intracellular Na+ content or Na+/K+ ratio. In EDL muscle, the ouabain-suppressible 86Rb+ uptake was stimulated by only 58%, whereas intracellular Na+ content and Na+/K+ ratio were increased by around 70%. Na(+)-loading of the muscles by exposure to K(+)-free or K(+)-Ca(2-)-Mg(2+)-free buffer stimulated the ouabain-suppressible 86Rb+ uptake in the two muscles to roughly the same extent, but in EDL muscle this was associated with a more than twofold larger increase in Na+/K+ ratio. When the Na+ influx was increased by exposure to veratridine similar results were obtained. Graded variation in intracellular Na+ content was achieved by exposure to monensin. In soleus muscle, a 25% increase in intracellular Na+/K+ ratio resulted in a doubling of the ouabain-suppressible 86Rb+ uptake, whereas a doubling of the Na(+)-K+ transport rate in EDL muscle required a 140% increase in Na+/K+ ratio. The results indicate that in soleus muscle the Na+/K+ pump is much more sensitive to changes in intracellular Na+ content than in EDL muscle. This might explain the larger activation of the Na(+)-K+ pump in slow-twitch muscle during electrical stimulation and might be of significance for the activation of the Na(+)-K+ pump in vivo during work.