The distribution of substance P and leucine enkephalin in mid- and fore-brain areas of the rat was studied using a radioimmunocytochemical method. The secondary antibody was labeled with 125I and the sections apposed to LKB Ultrofilm or emulsion-dipped. In alternate sections an extensive distribution of substance P and enkephalin immunoreactive material was seen in frontal, cingulate, retrosplenial, and entorhinal cortices. Substance P and enkephalin exhibited a remarkable overlap in many of these cortical areas as well as in the nucleus accumbens, caudate, portions of the hypothalamus, amygdala, thalamus and central gray. Differences in distribution were seen in the retrosplenial cortex, septum, ventromedial hypothalamus, hippocampus, the substantia nigra and the superior colliculus. The results provide a detailed immunohistochemical demonstration of the laminar patterns of substance P and enkephalin in the cortex of the rat. The results are discussed in terms of the interaction of substance P and enkephalin. The matches and mismatches of immunoreactive substance P and enkephalin and the locations of their receptors are also examined.