Projections from the prefrontal cortex to subdivisions of the dorsal raphe nucleus were investigated in the rat using retrograde and anterograde tracing methods. A retrograde tracer, gold-conjugated horseradish peroxidase (WGA-apo-HRP-gold), was injected into each subdivision of the dorsal raphe including lateral wing, dorsomedial, and ventromedial areas. The majority of retrogradely labeled cells were located in the prelimbic, infralim-bic, and dorsal peduncular areas of the medial prefrontal cortex. A few cells were also identified in the cingulate, various regions of the orbital, and agranular insular cortices. Secondly, an anterograde tracer, Phaseolus vulgaris leucoagglutinin (PHA-L), was injected into the medial prefrontal cortex involving the prelimbic or infralimbic areas. Axonal fibers with varicosities were identified in all subdivisions of the DR including the lateral wing, dorsomedial, and ventromedial areas. Projections were bilateral, with ipsilateral predominance. Axonal fibers were observed at the lateral border of medial longitudinal fasciculus or in the interfascicular region at the midline. The present findings demonstrate that both the midline and lateral wing regions of the dorsal raphe nucleus receive excitatory input from cognitive and emotional centers of the cerebral cortex.