In order to investigate the mechanism of Na and K transport, rabbit cortical collecting tubules were perfused in vitro and the concentrations of Na and K in lumen and bathing fluid and the transtubular electrical potential difference (PD) were measured. When the perfusate and external bath contained 150 Na-5 K (mEq liter(-1)), the sodium concentration decreased and the potassium concentration increased by an approximately equal amount in collected tubular fluid. The transtubular electrical potential was equal at both ends of the tubule in the steady state and ranged between 21 and 67 mv, lumen negative. In all tubules perfused at rates less than 0.5 nl min(-1), the K concentration of the collected fluid was higher and the Na concentration lower than that predicted for electrochemical equilibrium between lumen fluid and external bath, evidence for active transtubular transport of both cations. These results differ from those observed in rat distal tubule in which potassium secretion is passive. Active Na and K transport and the transtubular PD were decreased by (a) ouabain, (b) removal of sodium from the perfusate, or (c) removal of potassium from the external bath, evidence of interdependence of Na and K transport. The dependence of active K secretion on intraluminal Na concentration accounts for the phenomenon of "distal" Na-K exchange noted previously in clearance and stop-flow studies. The mechanism of Na transport may in part be electrogenic since the rate of decline of the transtubular PD in low K media was faster than could be accounted for on the basis of a reduction in cell potassium concentration.