Subregional distributions of serotonin1A receptors and serotonin transporters within the human dorsal raphe nucleus (DR) were determined by quantitative autoradiographic analyses of radioligand binding in tissue sections. [3H]8-Hydroxy-2-(di-n-propyl)aminotetralin (8-OH-DPAT) and [3H]paroxetine were used to label, respectively, serotonin1A receptors and serotonin transporters in the subnuclei of the DR, which were delineated on the basis of tryptophan hydroxylase (TrpOH) immunoreactivity. [3H]8-OH-DPAT binding was coextensive with the TrpOH-immunoreactive cell bodies and fibers but was distributed unevenly among the subnuclei. In contrast, [3H]paroxetine binding was present throughout the central gray matter, with relatively homogeneous labeling across the subnuclei of the DR. In rostral sections, [3H]8-OH-DPAT binding (fmol/mg protein) in the dorsal subnucleus was lower than that in the ventral or the interfascicular subnucleus. Within the interfascicular subnucleus, [3H]8-OH-DPAT binding decreased progressively in a rostral-to-caudal fashion. The highest levels of [3H]8-OH-DPAT binding were found in the ventrolateral subnucleus at the level of the caudal extent of the trochlear nucleus. The influence of age and postmortem interval on radioligand binding was also examined. These data in the human DR indicate that serotonin1A receptors are differentially distributed among the subnuclei and along the rostro-caudal axis of the midbrain raphe, and serotonin transporters appear to be relatively evenly distributed throughout the DR. Subregional analyses of such serotonergic markers may prove useful in evaluating the role that serotonin may play in depression, schizophrenia, and suicide.