The adrenal gland of mammals contains high concentrations of peripheral-type benzodiazepine receptors (PBR) and diazepam-binding inhibitor (DBI), a polypeptide which acts as an endogenous ligand for PBR. The aim of the present study was to investigate the localization of DBI and PBR in the adrenal gland of the frog Rana ridibunda. Reverse transcription followed by polymerase chain reaction with specific primers for the frog DBI cDNA showed the presence of DBI mRNA in frog adrenal gland extracts. The cellular distribution of DBI and PBR was investigated using an antiserum against the octadecaneuropeptide DBI [33-50] (ODN) and antibodies against the 18-kDa isoquinoline binding protein subunit of PBR (IBP), respectively. ODN-like immunoreactivity was found in chromaffin cells and in Stilling s cells, but not in adrenocortical cells. IBP-like immunoreactivity was observed in chromaffin cells, in Stilling s cells and in a small proportion (11%) of steroid-secreting cells. The ODN- and IBP-immunoreactive materials were homogeneously distributed in the cytoplasm of chromaffin cells and concentrated at the periphery of large cytoplasmic vesicles in Stilling s cells. The proportion of ODN-positive Stilling s cells showed marked circannual variations with a maximum in July. Similarly, the proportion of IBP-positive Stilling s cells was 17 times higher in July than in December. These results indicate that, in the frog adrenal gland, DBI-related peptides and PBR are simultaneously expressed in chromaffin cells and Stilling s cells, suggesting that endogenous ligands for PBR may play a physiological role in the control of adrenal cell activity.