Aminoacylhistidine dipeptides are present in the nervous tissue of many species. The olfactory mucosa and bulb of many vertebrates are rich in carnosine (beta-alanyl-L-histidine). Two related dipeptides homocarnosine (gamma-aminobutyryl-L-histidine) and anserine (beta-alanyl-N-methyl-L-histidine) are present in the CNS of mammals and birds, respectively. This manuscript describes the production, characterization and use in immunolocalization studies of antisera directed against carnosine and anserine. The anserine antiserum is highly specific for anserine while the carnosine antiserum cross-reacts with all three dipeptides. The differential specificity of the antisera, coupled with chemical characterization of the dipeptide composition of various brain regions, has permitted assignment of the cellular localization of the various dipeptides. Immunocytochemical localization of anserine has not been previously reported. Carnosine immunoreactivity in the olfactory system is restricted to the mature neurons in the olfactory mucosa, their axons and synaptic terminations in the glomerular layer of the olfactory bulb. Similar reactivity is seen in the accessory olfactory system. Astrocytes and cerebellar Bergmann glia seem to account for all the non-olfactory carnosine-like immunoreactive staining in the rodent brain. In contrast, in the avian CNS where anserine is chemically abundant, anserine-like immunoreactivity is widespread and apparently exclusively associated with glial cells. Thus, the olfactory receptor neurons appear to be the only neuronal population that expresses carnosine. Elsewhere in the CNS the aminoacylhistidine dipeptides are associated with various populations of glia.