The two neuronal tracers, Phaseolus vulgaris leucoagglutinin (PHA-L) and Cholera toxin subunit B (CHB) were used in order to study a possible neuronal projection from the entopeduncular nucleus to the ventral mesencephalon in the rat. Both tracers were identified in brain sections by means of immunohistochemistry. After injection of PHA-L in the entopeduncular nucleus PHA-L-positive nerve fibers observed in the mesencephalon were moderate in number and mostly restricted to the mediodorsal part of the substantia nigra, pars reticulata. In addition, a low number of PHA-L-immunoreactive nerve fibers was found in the substantia nigra, pars compacta, the rostral part of the ventral tegmental area, and in the deep mesencephalic nucleus. In agreement with these observations, several labeled neurons were observed in the ipsilateral entopeduncular nucleus after injections of CHB in the substantia nigra. These results indicate the presence of a direct neuronal projection between the two major output channels of the basal ganglia.