We have recently demonstrated that the avian brain as well as the peripheral steroidogenic glands produces pregnenolone, the main precursor of steroid hormones, on the basis of biochemical studies. Therefore, the immunohistochemical and Western immunoblotting analyses with a polyclonal antibody directed against the purified bovine adrenal cytochrome P450scc, which is involved in pregnenolone formation, was undertaken to investigate the localization of a cytochrome P450scc-like substance in the brain of adult male Japanese quails. P450scc-like immunoreactive cells were distributed in several telencephalic, diencephalic and mesencephalic regions. An intense immunoreaction was observed in somata of Purkinje cells and in the dendrites extending through the cerebellar molecular layer. Clusters of immunoreactive cell bodies were also detected in the hyperstriatum accessorium (HA), the ventral portions of both the archistriatum and the corticoid area, the preoptic area (POA), the anterior hypothalamus (AHy) and the dorsolateral thalamus (DL). Preadsorbing the antibody with cytochrome P450scc resulted in a complete absence of P450scc-like immunoreactivity in all of positively stained cells. On Western immunoblotting with the P450scc antibody, a P450scc-like substance was present in homogenates of the several brain regions that contain immunohistochemically stained cells. Immunohistochemical experiments using both antibodies against P450scc and the glial fibrillary acidic protein (GFA), a specific marker of glial cells, indicated that many HA cells contained both P450scc-like and GFA-like immunoreactivities. However, no immunoreactivity for GFA was observed in Purkinje cells and the cells localized in the ventral portion of the corticoid area, despite the presence of P450scc-like immunoreactivity. These results confirm our previous findings of pregnenolone biosynthesis and suggest the presence of a P450scc-like substance in both neuronal and glial cells of the quail brain.