Abstract A 41-residue urotensin I neuropeptide (H-UI) was isolated from urophyses of the marine teleost Hippoglossoides elassodon (the flathead sole). The peptide was recognized by its partial cross-reactivity in a radioimmunoassay developed for Catostomus (sucker) Ul (S-UI), and was purified by reversed-phase high-performance liquid chromatography. The amino-acid sequence was shown to be H-Ser-Glu-Glu-Pro-Pro-Met-Ser-lle-Asp-Leu-Thr-Phe-His-Met-Leu-Arg-Asn-Met-lle-His-Arg-Ala-Lys-Met-Glu-Gly-Glu-Arg-Glu-Gln-Ala-Leu-lle-Asn-Arg-Asn-Leu-Leu-Asp-Glu-Val-NH(2). H-UI is 66% homologous with S-UI and 63% homologous with Cyprinus (carp) Ul (C-UI). Like S- and C-UI, H-UI is about 50% homologous with the frog skin peptide sauvagine and with Catostomus and mammalian corticotropin-releasing factors. H-UI had similar vasodilatory effects in mammals, and similar adrenocorticotropin-releasing effects (in rat and goldfish) to S-UI, C-UI, sauvagine and the corticotropin-releasing factors, but had relatively low potency (e.g. 10% to 30% of the vasodilatory potency of S- and C-UI) in all the bioassay systems studied.