Distribution of [S-35]-TBPS binding sites was studied in various structures of brain in mouse and guinea pig and in cortex of monkey and in hippocampus of postmortem human brain. As it is observed for rat brain, high densities of [S-35]-TBPS binding sites were found in layer IV of cortex in the four species, and in thalamus of mouse and guinea pig. Intermediate densities of binding sites were observed in superficial and deep layers of cortex in those four species and in hippocampus of mouse, guinea pig, and human. In all brain structures studied, 5alpha3alphaP and picrotoxin produced a dose-dependant inhibition of [S-35]-TBPS binding. No significant interregion or interspecies differences could not be detected for IC50 values of 5alpha3alphaP or picrotoxin to inhibit [S-35]-TBPS from its binding sites. In all regions studied, IC50 values were close to 1.5 X 10(-6) M for 5alpha3alphaP and 2.3 X 10(-7) M for picrotoxin.