The neostriatum and its major afferent transmitters, dopamine and glutamate, play a critical role in behavior, but relatively little information is available on their postsynaptic effects in behaving animals. As a first step in addressing this shortcoming, single-unit electrophysiology was combined with iontophoresis in the neostriatum of awake, unrestrained rats. Relative to periods of quiet rest, most neurons (58 of 77) changed discharge rate in close temporal association with movement, while the remainder showed no such relationship. When animals resumed a resting posture, iontophoretic current-response curves were established for dopamine and glutamate as well as for ascorbate, a modulator of neostriatal function released from glutamatergic terminals. Application of either glutamate or ascorbate produced current-dependent increases in activity in all neurons, although this effect was somewhat less pronounced for nonmotor cells. In both types of neurons, the excitatory effect of ascorbate either diminished or shifted to an inhibition at high ejection currents. Dopamine, on the other hand, routinely excited motor-related, but inhibited nonmotor-related neurons. Further assessment of motor-related neurons revealed that in most cases the excitatory effects of either glutamate or dopamine alone were supra-additive when these compounds were either administered together or co-administered with ascorbate. Our results suggest that the response of neostriatal neurons to glutamate or dopamine depends, at least in part, on the motor responsiveness of these cells. Motor-related neurons, moreover, respond to the co-administration of glutamate and dopamine with synergistic increases in firing rate. Ascorbate also influences neostriatal activity, but the postsynaptic action of this substance cannot be explained as a simple interaction with either glutamatergic or dopaminergic mechanisms.