Ovine genomic urocortin: cloning, pharmacologic characterization, and distribution of central mRNA.
Urocortin (Ucn), the newest member of the corticotropin-releasing factor (CRF) family of peptides, has been demonstrated to have significant physiologic and behavioral effects following its peripheral and central administration, respectively. In order to assess the differences in Ucn across species, an 18-kb sheep genomic DNA fragment encoding urocortin was isolated by the hybridization screening of a lambda phage library with a probe generated from rat urocortin (rUcn) cDNA. The sheep clone contains a region that is 84% and 88% homologous to the coding region of rUcn and human Ucn (hUcn), respectively and encodes an ovine Ucn (oUcn) that is predicted to be identical to the rat peptide. Competitive binding assays demonstrated oUcn to have a high affinity (Ki=0.1 nM) for the sheep CRF-binding protein (CRF-BP) and localization studies by in situ hybridization have shown that the distribution of oUcn messenger RNA in sheep brain shares with that of rUcn in rat brain a predominant locus of expression in the Edinger-Westphal nucleus of the midbrain, though some secondary sites of expression reported in rat are not conserved. These findings demonstrate that, even across diverse species, Ucn is highly conserved with respect to its structure and pharmacology unlike CRF where significant amino acid substitutions between the rat/human and sheep peptides may underlie differences in neuroendocrine regulation.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001923:Brain Chemistry / Q000502:physiology*; D003001:Cloning, Molecular; D003346:Corticotropin-Releasing Hormone / Q000235:genetics*; D016678:Genome; D006801:Humans; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D011485:Protein Binding; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D012756:Sheep; D013045:Species Specificity; D054832:Urocortins