Characterization of four Mx isoforms in the European eel,Anguilla anguilla

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Abstract

Mx protein is known to play an important role in vertebrate immune response to viral infection. In this study, cDNA sequences of four Mx isoforms, designated as MxA, B, C and D were characterized in the European eel, Anguilla anguilla. These sequences contained an open reading frame of 1899, 1896, 1866, 1779 bp, flanked by 95, 53, 138, 69 bp of 5′ untranslated region and 389, 241, 136, 124 bp of 3′ untranslated region, respectively. A phylogenetic tree constructed with Mx peptide sequences from vertebrates revealed that MxA, C and D in the European eel formed into a clade containing zebrafish MxA and MxB and Mx proteins in other teleosts, whereas MxB in the eel was clustered together with zebrafish MxD, MxG and MxF. The transcription level of all Mx isoforms increased in a poly I:C dose-dependent manner in peripheral blood leukocytes of eels, as revealed by real-time PCR. A further experiment was conducted to reveal the temporal change in expression of these isoforms in various organs/tissues following poly I:C stimulation, and significant increase in expression was observed at various degrees in different organs or in different sampling occasions within the 12 h experimental period. In particular, MxA had the highest level of increase, while MxB had the lowest; and three isoforms, MxA, MxB and MxD had the highest increase in intestine, while the highest increase of MxC expression was observed in liver. These four isoforms of eel Mx are thus expressed differentially, and further work is certainly required to clarify the activity of promoter elements and antiviral activity of these Mx isoforms.

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