Spleen tyrosine kinase from Nile tilapia (Oreochromis niloticus): Molecular characterization, expression pattern upon bacterial infection and the potential role in BCR signaling and inflammatory response

摘要

Spleen tyrosine kinase (SYK), a member of non-receptor tyrosine kinase family, plays an important role in immune responses against pathogen infection, which is capable of activating B cells signaling pathway and regulating inflammatory response. In this study, Nile tilapia (Oreochromis niloticus) ortholog (OnSYK) was identified and characterized at expression pattern against bacterial infection, function in B cells activation pathway and inflammatory response. The cDNA of OnSYK ORF contained 1851 bp of nucleotide sequence encoding polypeptides of 616 amino acids. The deduced OnSYK protein was highly homologous to other species SYK, containing two SH2 domains and a TyrKc domain. Spatial mRNA expression analysis revealed that OnSYK had wide tissue distribution and was highly expressed in the liver. After challenge of Streptococcus agalactiae (S. agalactiae) in vivo, mRNA expression of OnSYK was significantly up-regulated in the head kidney, spleen and liver. The up-regulation of OnSYK transcript was also displayed in the head kidney and spleen leukocytes stimulation with S. agalactiae and LPS in vitro, which was confirmed at protein level in the head kidney leukocytes by FACS analysis. In addition, after induction with mouse anti-OnIgM monoclonal antibody in vitro, the expressions of OnSYK and its downstream molecules (OnLYN, OnBLNK and OnAP-1) were significantly up-regulated in the head kidney leukocytes, and pharmacological inhibition of SYK activity with inhibitor (P505-15) significantly attenuated the expressions of OnLYN, OnBLNK and OnAP-1. Moreover, upon LPS challenge, the expressions of OnSYK, OnTNF-alpha, OnIL-6 and OnAP-1 were also up-regulated in the head kidney monocytes/macrophages. After treatment with SYK inhibitor (BAY 61-3606), the expressions of OnTNF-alpha, OnIL-6 and OnAP1 were inhibited in the LPS-challenged head kidney monocytes/macrophages. Taken together, the results of this study indicated that OnSYK, playing potential roles in BCR signaling and inflammatory response, was likely to get involved in host defense against bacterial infection in Nile tilapia.