The cytological and topographical distribution of S-100 and glial fibrillary acidic (GFA) proteins in the adult rat brain has been compared using the horseradish peroxidase-labelled antibody technique. Both proteins are present in astrocytes and structures composed of astrocytic processes, namely the glial limitans and the perivascular membranes, but the cytological localization varies between the two proteins. S-100 is found in the nucleus and cytoplasm whereas GFA protein is confined to the cytoplasm. Neither is found in neurons, but S-100 is present in some oligodendroglia, suggesting a general regulatory role in glia. Although GFA protein is present in both protoplasmic and fibrous astrocytes, it is more prominent in the latter, confirming its association with astrocytic filaments.