title: Molecular characterization of LEAP-2 cDNA in common carp (Cyprinus carpio L.) and the differential expression upon a Vibrio anguillarum stimulus; indications for a significant immune role in skin.
abstract: LEAP-2 is a cysteine-rich cationic antimicrobial peptide (AMP) playing an important role in host innate immune system. LEAP-2 genes have been identified from higher vertebrates and several fish species. Here we report the cloning and identification of two LEAP-2 cDNA sequences from the liver of common carp (Cyprinus carpio L.). The LEAP-2A cDNA was 1325 bp long and contained an ORF of 279 bp encoding a protein of 92 amino acids. The LEAP-2B cDNA was 608 bp long and contained an ORF of 276 bp encoding a protein of 91 amino acids. Both LEAP-2 proteins consisted of 41 amino acid residues and shared four cysteines at the conserved positions in the predicted mature peptides, highly similar to LEAP-2 of other species. Sequence alignment showed that LEAP-2 amino acid sequences were well conserved in different species, and the phylogenetic relation of LEAP-2 was coincident with evolution of biological species. Expression analysis data revealed that LEAP-2A and LEAP-2B mRNAs were expressed in a wide range of common carp tissues including liver, spleen, head kidney, skin, gills, hindgut and foregut. When injected intraperitoneally with Vibrio anguillarum, the expression level of common carp LEAP-2A was quickly up-regulated in liver, spleen, head kidney, skin, gills, foregut and hindgut, however, the expression level of LEAP-2B was similarly up-regulated in spleen, skin, gills and hindgut but not in liver, head kidney and foregut. Our results showed that the LEAP-2A had a markedly high constitutive expression in skin, and the LEAP-2A and the LEAP-2B had a significantly high up-regulated expression after stimulus in skin. This differential expression of LEAP-2 in common carp suggests that it may play a key role in immune responses against invading pathogens and both LEAP-2 molecules may be involved in mucosal immunity.
authors.affiliation: Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Science, Shandong Normal University, No. 88, East Wenhua Road, Jinan 250014, PR China; Cell Biology and Immunology Group, Department of Animal Sciences, Wageningen University, PO Box 338, 6700 AH Wageningen, The Netherlands.
authors.forename: Guiwen
authors.initials: G
authors.lastname: Yang
authors.affiliation: Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Science, Shandong Normal University, No. 88, East Wenhua Road, Jinan 250014, PR China.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Guo
authors.affiliation: Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Science, Shandong Normal University, No. 88, East Wenhua Road, Jinan 250014, PR China.
authors.forename: Hua
authors.initials: H
authors.lastname: Li
authors.affiliation: Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Science, Shandong Normal University, No. 88, East Wenhua Road, Jinan 250014, PR China.
authors.forename: Shijuan
authors.initials: S
authors.lastname: Shan
authors.affiliation: Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Science, Shandong Normal University, No. 88, East Wenhua Road, Jinan 250014, PR China.
authors.forename: Xueqing
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Cell Biology and Immunology Group, Department of Animal Sciences, Wageningen University, PO Box 338, 6700 AH Wageningen, The Netherlands. Electronic address: j.rombout@freeler.nl.
authors.forename: Jan H W M
authors.initials: JH
authors.lastname: Rombout
authors.affiliation: Shandong Provincial Key Laboratory of Animal Resistance Biology, College of Life Science, Shandong Normal University, No. 88, East Wenhua Road, Jinan 250014, PR China. Electronic address: an.liguo58@gmail.com.
authors.forename: Liguo
authors.initials: L
authors.lastname: An
mesh_terms: D000818:Animals; D001483:Base Sequence; D002347:Carps / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D020869:Gene Expression Profiling / Q000662:veterinary; D005786:Gene Expression Regulation / Q000276:immunology*; D064451:Hepcidins / Q000235:genetics*; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA / Q000662:veterinary; D012867:Skin / Q000276:immunology*; D013045:Species Specificity; D014733:Vibrio / Q000276:immunology*
chemical_list: D018076:DNA, Complementary; D064451:Hepcidins
keywords: Common carp (Cyprinus carpio L.); Innate immunity; LEAP-2; Molecular cloning; mRNA expression