We studied the interactions between glutamate and dopamine in the pars reticulata of the substantia nigra by using microdialysis in unanaesthetized rats. Increased extracellular levels of glutamate in the pars reticulata were obtained by microinjecting the muscarinic agonist carbachol into the ipsilateral subthalamic nucleus. The increase of glutamate levels was followed by increments in extracellular levels of dopamine and GABA. Increased levels of the three neurotransmitters were also observed during the administration of N-methyl-D-aspartate through the microdialysis probe. The increase in glutamate and GABA caused by N-methyl-D-aspartate was blocked by SCH 23390, a selective D1 antagonist. However, the D1 antagonist did not prevent the increase in dopamine levels. The selective D1 agonist SKF 38393, added to the microdialysis probe, increased the levels of the three neurotransmitters. However, after the lesion of the subthalamic nucleus with kainic acid, SKF 38393 increased only the level of GABA but not those of glutamate and dopamine. In addition, the lesion of the subthalamic nucleus produced a drastic (80%) fall in the extracellular levels of glutamate. These data suggest that glutamate, through N-methyl-d-aspartate receptors, stimulates the release of dopamine from dopaminergic dendrites present in the substantia nigra pars reticulata, and that dopamine in turn stimulates the release of glutamate and GABA. Both effects are mediated by D1 dopamine receptors present on subthalamonigral and striatonigral axon terminals, respectively.