A direct synaptic contact between nigrostriatal dopamine neurons and substance P axons in the substantia nigra was demonstrated using the immunoelectron microscopic mirror technique combined with the fluorescent double-staining method for transmitter-specific projections. Substance P-immunoreactive terminals were found to make synaptic contact with nigral cells exhibiting tyrosine hydroxylase immunoreactivity and retrograde fluorescent labeling following injection of biotinylated lectin into the neostriatum. It appears that substance P afferents directly affect nigrostriatal dopamine neurons in the substantia nigra via the synaptic contacts.