A NLRC3-like gene from blunt snout bream (Megalobrama amblycephala): Molecular characterization, expression and association with resistance to Aeromonas hydrophila infection. Issue 63 (April 2017)

• Record Type: Journal Article
• Authors: Zhou, Fengjuan; Zhan, Qifeng; Ding, Zhujin; Su, Lina; Fan, Jun; Cui, Lei; Chen, Nan; Wang, Weimin; Liu, Hong
• Is Part Of: Fish & shellfish immunology. Issue 63(2017)
• Page Start: 213
• Page End: 219
• Publication Date: 2017-04
• Subjects: MamNLRC3-like -- NLRC3-like -- High resolution melting -- Association analysis -- Susceptibility/resistance -- SNPs; Fishes -- Immunology -- Periodicals; Shellfish -- Immunology -- Periodicals; Poissons -- Immunologie -- Périodiques; Crustacés -- Immunologie -- Périodiques; 571.9617
• DOI: 10.1016/j.fsi.2017.02.018
• Languages: English
• ISSNs: 1050-4648

Abstract

Abstract: NLRC (the nucleotide-oligomerization domain (NOD)-like receptor subfamily C) consists of teleost-specific NLRs (NOD-like receptors) and plays pivotal roles in microbial recognition and regulation of innate immune response. In this study, we cloned and characterized a NLRC3-like gene ( MamNLRC3-like ) from blunt snout bream ( Megalobrama amblycephala ) by using the quantitative real-time PCR method, and analyzed the correlation between its polymorphisms and resistance to Aeromonas hydrophila infection. The full length cDNA of MamNLRC3-like was 2863 bp, with a 5′-UTR of 169 bp, ORF of 2301 bp and 3′-UTR of 393 bp, encoding 766 amino acid residues. MamNLRC3-like consisted of a conserved NACHT domain, a fish-specific NACHT associated domain (FISNA) and a PYRIN effective domain. During early developmental stages, MamNLRC3-like was most highly expressed at 39.4 hpf (hours post fertilization, hatching stage) and constitutively detected in various healthy tissues. The expression of MamNLRC3-like was strongly induced by A. hydrophila infection and peaked in the head kidney, liver, intestine, and trunk kidney at 12 h post infection. Six SNPs (single nucleotide polymorphisms) from MamNLRC3-like, with 2 in introns and 4 in exons, were identified by direct sequencing, in which 6515C/T was a novel non-synonymous mutation. HRM (high resolution melting) genotyping of the 6 loci showed that locus 6515C/T SNP was associated with the resistance/susceptibility of blunt snout bream to A. hydrophila infection ( P < 0.01). The CC genotype fish were more resistant than CT carriers ( P < 0.01). This study suggests that the MamNLRC3-like gene might play an important role in the innate immunity of blunt snout bream and could be used as a candidate marker for genetic selection of A. hydrophila -resistant blunt snout bream. Highlights: MamNLRC3-like was cloned and characterized. Its expression in tissues, early development and bacterial infection was examined. Six SNPs in MamNLRC3-like were identified. Locus 6515C/T was associated with resistance to bacterial septicemia.