In vivo microdialysis and single-cell extracellular recordings were used to assess the involvement of serotonin(4) (5-HT(4)) receptors in the effects induced by morphine, amphetamine and cocaine on nigrostriatal and mesoaccumbal dopaminergic (DA) pathway activity. The increase in striatal DA release induced by morphine (2.5 mg/kg, s.c.) was significantly reduced by the selective 5-HT(4) antagonists GR 125487 (0.1 and 1 mg/kg, i.p.) or SB 204070 (1 mg/kg, i.p.), and potentiated by the 5-HT(4) agonist prucalopride (5 mg/kg, i.p.). Neither of these compounds affected morphine-stimulated DA release in the nucleus accumbens. In both regions, amphetamine (2 mg/kg, i.p.) and cocaine (15 mg/kg, i.p.) induced DA release was affected neither by GR 125487 nor by prucalopride. None of the 5-HT agents used modified basal DA release in either brain region. Finally, GR 125487 (445 microg/kg, i.v.), whilst not affecting basal firing of DA neurons within either the substantia nigra pars compacta nor the ventral tegmental area, significantly reduced morphine (0.1-10 mg/kg, i.v.) stimulated firing of nigrostriatal DA neurons only. These results confirm that 5-HT(4) receptors exert a state-dependent facilitatory control restricted to the nigrostriatal DA pathway, and indicate that 5-HT(4) receptors selectively modulate DA exocytosis associated with increased DA neuron firing rate.