Corticotropin Releasing Factor (CRF) immunoreactivity was localized within the rat central nervous system with immunocytochemical methods, using a highly specific anti-serum to rat-CRF. In adult rats abundant CRF immunoreactivity (CRF-ir) was found in perikarya of the inferior olivary nucleus, in climbing fibres in the molecular layer, and mossy fibres in the granular layer of the cerebellum and in the cerebellar nuclei. CRF-ir climbing fibres were found in a longitudinal zonal pattern throughout the molecular layer. In developing cerebellum only a few CRF-positive fibres were seen on postnatal day 8. From day 8 onwards a gradual increase in CRF-ir was found in both the molecular and granular layer with formation of CRF-positive nests around Purkinje cellbodies starting at day 12. Subsequent outgrowth into the molecular layer could be followed, leading to an adult level of CRF-ir in the cerebellum at approximately day 22 postnatally. CRF-ir in mossy fibres has as adult appearance from approximately day 18. The temporal and morphological changes of CRF-ir in fibres in the cerebellum of immature rats followed closely the morphological development of climbing and mossy fibres. The present findings indicate that CRF (or related substances) are already present in both climbing and mossy fibres during their outgrowth into the cerebellum.