Our current understanding of the corticotropin-releasing factor (CRF) system distribution in the teleost brain is restricted by limited immunohistochemical studies and a lack of complete transcriptional distribution maps. The present study used in situ hybridization to localize and compare CRF, urotensin I (UI), and CRF-binding protein (CRF-BP) expression in the brain of adult zebrafish (Danio rerio). All three peptides were localized in the preoptic area, periventricular hypothalamic and tectal regions, and dorsal part of the trigeminal motor nucleus. CRF and UI were both expressed in several nuclei of the dorsal telencephalon, whereas CRF and CRF-BP were both expressed in the ventral nucleus of the ventral telencephalon. Sole expression of CRF and CRF-BP was apparent in the olfactory bulbs and superior raphe nucleus, respectively, whereas only UI was observed in the corpus mamillare, nucleus of the medial longitudinal fascicle, dorsal tegmental nucleus, nucleus lateralis valvulae, and nucleus interpeduncularis. A major finding of this study was the general regional overlapping of CRF-BP with its ligands and a tendency to be expressed in tandem with CRF rather than UI. Overall, the mRNA expression patterns outlined in this study support the stress-related neuroendocrine, autonomic, and behavioral functions generally ascribed to the vertebrate CRF system and suggest some unique functional roles for CRF and UI in the teleost brain. J. Comp. Neurol. 502:783–793, 2007. © 2007 Wiley-Liss, Inc.