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The Characterization of Ayu (Plecoglossus altivelis) akirin1 Gene and Regulation of Function in Monocytes Macrophages |
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Abstract Akirin, a conserved nuclear factor, is associating with immune functions in animal via regulating cytokine transcription and immune cell chemotaxis. Here, an akirin1 cDNA sequence (Paakirin1) from ayu (Plecoglossus altivelis) was cloned. The Paakirin1 was 1817 bp in length encoded a 182 aa polypeptide with a molecular weight of 20.68 kD(GenBank accession No. KP278589). Sequence comparisons for similarity showed that Paakirin1 was highly similar to akirin1 in fish, such as, rainbow trout (Oncorhynchus mykiss) akirin1 (86%), followed by Arctic char (Salvelinus alpinus) and Atlantic salmon (Salmo salar) akirin1 (85%). Multiple sequence alignment analysis showed that Paakirin1 possessed nuclear localization signal. Phylogenetic analysis showed that fish akirin1 genes grouped tightly into one big cluster. Paakirin1 clustered together with the akirin1s of salmonidae family. Semi-quantitative PCR data showed that Paakirin1 was mainly expressed in head-kidney, spleen, intestine, and monocytes macrophages. After Vibrio anguillarum infection, Paakirin1 mRNA was up-regulated in head-kidney and spleen, while down-regulated in intestine. Furthermore, Paakirin1 mRNA was also up-regulated in monocytes macrophages after lipopolysaccharide (LPS) or polyinosinic-polycytidylic acid (PolyI:C) treatment. After Paakirin1 siRNA treatment in monocytes macrophages, the mRNA expression of interleukin-1β (IL-1β) and tumor necrosis factor (TNF) induced by V. anguillarum was unaltered, while its chemotaxis toward to recombinant CXC chemokine ligand 8 (CXCL8) protein was down-regulated. In summary, Paakirin1 alters its expression after infection and mediates chemotaxis of ayu monocytes macrophages. The research provides a theoretical basis and reference for the further investigation of function and regulation of monocytes macrophages in fish.
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Received: 26 March 2015
Published: 24 July 2015
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