Similar to other dopaminergic systems, the dopaminergic neurons innervating the stalk-median eminence (SME) and posterior pituitary (PP) possess an uptake mechanism for dopamine (DA). However, the extent of DA reuptake in these tissues and its physiological significance are debated since much of the released DA is removed by the hypophysial portal vasculature before recapture. The objectives of this study were: (1) to establish in vitro conditions for examining the effects of reuptake inhibitors on DA release from the PP and SME; (2) to compare the effects of nomifensine, diclofensine and amphetamine on DA release from the SME and PP; and (3) to distinguish between reuptake and releasing properties of these drugs. Individual SME and PP were dissected from ovariectomized rats and incubated in either a static or perifusion system. Media DA was extracted with alumina and quantitated by high performance liquid chromatography with electrochemical detection. The reuptake inhibitors, nomifensine, dichlofensine and amphetamine, in the presence of pargyline, a monoamine oxidase inhibitor, stimulated both basal and K(+)-evoked release of DA from the SME and PP under static incubation conditions. The drugs elicited a 2-3-fold higher increase in basal DA release from the SME as compared to the PP. Only amphetamine stimulated DA release in the perifusion system whereas nomifensine and diclofensine were without effects. We concluded that: (1) a mechanism for the reuptake of DA is operable in both the SME and PP; (2) the reuptake of DA appears to be more active in the SME than the PP; and (3) unlike amphetamine, nomifensine and diclofensine are pure reuptake inhibitors devoid of direct DA releasing activities.