Mechanisms of tetraethylammonium (TEA) transport were investigated in basolateral membrane (BLM) vesicles from rabbit renal cortex. Preloading vesicles with 10 mM TEA or 1 mM mepiperphenidol stimulated the 15-s uptake of [14C]-TEA compared with control vesicles (258 and 200%, respectively) and produced an overshoot of the equilibrium value (3 and 1.6 times, respectively). In the presence of a K+ gradient, net TEA uptake was also increased (and showed an overshoot of 2-fold) when the membrane potential of vesicles was made interior negative by adding valinomycin. Both TEA-TEA exchange and the voltage-driven net TEA transport were cis-inhibited by other organic cations, and a similar affinity order was found for both transport mechanisms (quinine greater than amiloride greater than morphine greater than mepiperphenidol greater than choline = N1-methylnicotinamide). This data suggests that the same transport protein might be responsible for both phenomena. Other experiments determined that the BLM vesicles had no TEA-H+ exchange, and that contamination of the vesicle population by brush-border membranes was negligible in terms of their contribution to TEA transport. These results demonstrate the presence of an exchange reaction for TEA in the rabbit renal BLM and thus imply carrier-mediated transport of TEA by these membranes.