In this study, we demonstrate that 6-hydroxydopamine (6-OHDA) can be used to produce a lesion of dopamine (DA) terminals in medial prefrontal cortex (mPFC) while sparing the noradrenergic innervation in this region. Furthermore, we determined the impact of these lesions on both extracellular DA in neostriatum, using in vivo microdialysis, and locomotor activity. Our results demonstrate that, whereas higher doses of 6-OHDA (> or = 4 micrograms) depleted both DA and norepinephrine (NE) in mPFC, 1 micrograms 6-OHDA produced a depletion of DA (-79%) without significantly affecting NE content (-13%). Selective depletion of DA content in mPFC did not alter basal levels of extracellular DA in neostriatum determined 14 days after the lesion. The lesion also did not alter the ability of acute tail pressure (30 min) to increase extracellular DA in neostriatum or to stimulate locomotor activity. Depletion of DA in mPFC did not alter the ability of d-amphetamine (1.5 mg/kg, i.p.) to increase intracellular DA in neostriatum. In contrast, the maximum amphetamine-induced increase in locomotor activity was attenuated in lesioned rats as compared with control rats (670 and 280 locomotor counts/15 min, respectively). These data suggest that in the intact system, DA terminals in mPFC do not regulate extracellular DA in neostriatum. In addition, these data confirm that DA terminals in mPFC can influence stimulant-induced locomotion.