Although ample evidence implicates the dopamine (DA) projection to the neostriatum and nucleus accumbens in motor and motivational processes, relatively little information is available on how DA alters neostriatal or accumbal functions under naturally occurring behavioral conditions. Further insight into neuron-behavior relationships can be achieved with the application of single-unit recording techniques, including iontophoresis and fast-scan cyclic voltammetry (FSCV), to awake, unrestrained animals. Single-unit recording has revealed that amphetamine, a widely abused psychomotor stimulant, activates motor-, but inhibits nonmotor-related neurons in neostriatum and nucleus accumbens. Although either response can be blocked by DA receptor antagonists, the amphetamine-induced activation also depends on an intact corticostriatal system, suggesting a role for glutamate (GLU). Both neostriatal and accumbal neurons are sensitive to iontophoretic application of either DA or GLU, but when applied during low-dose application of DA, the GLU signal is enhanced relative to background activity. In effect, DA appears to modulate GLU by strengthening the GLU signal-to-noise ratio. To assess DA release under behaviorally relevant conditions, FSCV has been used to obtain real-time measurements of DA efflux in a free-choice novelty test. DA efflux increased only during the brief period of entry into novelty, and the increase was confined to accumbal shell and the shell-core transition zone, the so-called shore. Neither accumbal core nor the overlying neostriatum showed a novelty-related DA change. Thus, DA release during behavior is not uniform and in the case of novelty appears targeted to the limbic-related area of accumbal shell. Further application of these and other in vivo technologies to ambulant animals is required to identify the complex mechanisms underlying both the release of DA and its effect on neostriatal and accumbal neurons during behavior.