The aim of this study was to investigate the distribution of serotonin (5-HT) receptors of type 6 (5-HT₆) in postmortem human prefrontal cortex, striatum and hippocampus. The brain samples were obtained from 6 subjects who had died for causes not involving primarily or secondarily the CNS. The 5-HT₆ receptor distribution was explored by the [¹²⁵I]SB-258585 binding to brain membranes followed by the pharmacological characterization, where possible, and by autoradiographic, immunohistochemical and immunofluorescence evaluations. A specific and saturable [¹²⁵I]SB-258585 binding was detected in striatum only, with a pharmacological characterization consistent with that of a 5-HT₆ receptor. The autoradiography showed the presence of a specific [¹²⁵I]SB-258585 binding distributed homogeneously in caudate, putamen and accumbens. The immunohistochemistry, carried out in the striatum only, coupled with the immunofluorescence with glial fibrillary acidic protein (GFAP) and parvalbumin (PV) showed the co-localization of 5-HT₆ receptor with PV, while indicating that this receptor subtype was expressed in neurons and not in astrocytes. Taken together, the present findings showed the presence of a higher density of 5-HT₆ receptors, as labeled by [¹²⁵I]SB-258585, in striatum than in hippocampus and prefrontal cortex, and specifically within the neuronal body. In addition, they would suggest that striatum is one of the major potential CNS targets linked to 5-HT₆ receptor modulation.