Whether Na+ movement through the plasma membrane plays a role in thyroid hormone uptake was investigated in intact rat soleus muscles. After preincubation for 120 min at 37 degrees C in modified Krebs-Ringer bicarbonate containing 140 or 5 mM Na+ plus choline or lithium to maintain osmolarity, muscles were incubated with 50 pM [125I]triiodo-L-thyronine (T3) or [125I]L-thyroxine (T4) for 60 min. T3 uptake was decreased when extracellular Na+ was replaced by either choline or lithium, the amount of decrease corresponding to the specific (or saturable) uptake component. Monensin, an ionophore that stimulates Na+ entry, increased T3 uptake at 140 mM Na+ but not at 5 mM Na+. Amiloride, a Na+/H+ exchange inhibitor, had no effect on T3 uptake under basal conditions or when Na+ was replaced by choline, but reversed the action of lithium. Ouabain, an inhibitor of Na+/K+ ATPase, reduced specific T3 uptake. T4 uptake was unaffected by low extracellular Na+. These results are consistent with a major role of Na+ movement in T3 uptake by skeletal muscle, but not in T4 uptake, and suggest an involvement of membrane pumps in this process.