Relatively high concentrations of adrenocorticotropic hormone (ACTH), beta-endorphin and alpha-melanocyte-stimulating hormone (alpha-MSH) were determined by radioimmunoassay in the nucleus of the solitary tract (NTS) of rats. Dense networks of immunoreactive fibers for these peptides were most prominent in the commissural part of the nucleus, where immunostained perikarya (8-15 per section) were also seen in colchicine-treated rats. Moderate peptide levels and moderately dense immunoreactive networks of these peptides were found in the lateral reticular nucleus (including the A1 and A5-C1 catecholaminergic cell groups) and the nucleus ambiguus. Ten different types of surgical lesions or transections were performed in the hypothalamus and the lower brainstem to determine the origin of ACTH, beta-endorphin and alpha-MSH in the brainstem baroreceptor centers. Except the commissural part of the NTS, the baroreceptor areas receive ACTH, beta-endorphin and alpha-MSH innervations from both the hypothalamic arcuate cells and local neurons in the NTS. Fibers in the commissural part of the NTS seem to be of local origin. Hypothalamic fibers to the rostral part of the NTS and the vasomotor A5-C1 cell groups descend in both a medial (through the periaqueductal central gray) and a lateral (ventrolateral tegmental fibers) pathway, whereas fibers to the caudal lateral reticular nucleus (A1 cell group) and the nucleus ambiguus may run only in the lateral pathway. The descending fibers may decussate somewhere in the caudal hypothalamus-rostral midbrain, but caudal to that level they run and terminate ipsilaterally. Fibers from the ACTH-, beta-endorphin- and alpha-MSH-containing cells in the NTS form a bundle arching between the NTS and the ventrolateral medulla and partially (40-55%) innervate the vasomotor and the vasodepressor areas, as well as the nucleus ambiguus.