title: Expression of an inducible nitric oxide synthase gene in rainbow trout Oncorhynchus mykiss.
abstract: Using oligonucleotide primers based on mammalian nitric oxide synthases (NOS), expression of an inducible NOS (iNOS) gene was detected in head kidney and gill tissue of bacterially-challenged rainbow trout. Three overlapping fragments were amplified by RT-PCR and used to construct a contiguous sequence of 1410bp, with high nucleotide homology to iNOS in birds (61%) and mammals (57-59%). The nucleotide sequence translated in one reading frame to produce a partial peptide containing 470 amino acids, with 69-71% amino acid homology with mammalian iNOS, 81% homology with chicken iNOS and 85% homology with a partial (492bp) goldfish iNOS sequence. In vitro stimulation of head kidney macrophages with LPS also induced expression of the trout iNOS RNA, with optimal expression seen using 20-50 microg/ml LPS at 2h to 6h post-stimulation. The evolutionary and functional significance of the trout iNOS sequence are discussed.
authors.affiliation: Department of Zoology and Medicine & Therapeutics, University of Aberdeen, UK.
authors.forename: K J
authors.initials: KJ
authors.lastname: Laing
authors.forename: L J
authors.initials: LJ
authors.lastname: Hardie
authors.forename: W
authors.initials: W
authors.lastname: Aartsen
authors.forename: P S
authors.initials: PS
authors.lastname: Grabowski
authors.forename: C J
authors.initials: CJ
authors.lastname: Secombes
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D015870:Gene Expression / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D019001:Nitric Oxide Synthase / Q000235:genetics*; D052247:Nitric Oxide Synthase Type II; D017686:Oncorhynchus mykiss / Q000235:genetics*; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms / Q000235:genetics
chemical_list: D017931:DNA Primers; D008070:Lipopolysaccharides; D020033:Protein Isoforms; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II