黄颡鱼甘露糖受体基因的克隆和功能分析
Cloning and functional analysis of mannose receptor from Chinese yellow catfish (Pelteobagrus fulvidraco)
摘要
Abstract
Mannose receptor (MR) is a member of C-lectin family. It is mainly expressed on the surface of macrophage and immature dendritic cells. It has been shown that MR not only plays a significant role in innate immune responses, but also initiates acquired immune responses through processing and presenting antigens to activate T cells. In this study, we cloned and characterizedpf MR of Chinese yellow catfish (Pelteobagrus fulvidraco). The expression profiles of pfMR mRNA in different tissues of P. fulvidraco were assessed by real-time quantitative reverse transcription PCR (qRT-PCR). To investigate the role of pfMR participating in the antibacterial infection, yellow catfish head kidney macrophages were infected with Edwardsiella ictaluri and competition binding test of mannan to MR was carried out. The phylogenetic tree analysis showed that MR of P. fulvidraco, Megalobrama amblycephala,Ctenopharyngodon idella,Danio rerio, and Oreochromis niloticus formed a clade. The mRNA of pfMR was expressed in all detected tissues with higher level in the kidney, spleen and muscle, and lowest expression in the blood. The expressions of pfMR,IL-1β and TNF-a mRNAs were all increased in cultured primary head-kidney macrophages after E. ictaluri infection. The infection of E. ictaluri also resulted in the over-expression of large number of superoxide anion () and nitric oxide (NO) in the macrophages. The infection of E. ictaluri could significantly activate the production of at 30 min post treatment, while the amount of NO in the macrophage was significantly elevated at 12 h post treatment. Mannan significantly inhibited the engulfed GFP-E. ictaluri by macrophages through competition binding to MR. The engulfed GFP-E. ictaluri was significantly reduced by addition of EDTA, and restored by addition of Ca2+. The present study indicated that MR in head kidney macrophages of yellow catfish mediated phagocytosis of E. ictaluri and it was Ca2+-dependent.关键词
黄颡鱼/甘露糖受体/鲖爱德华菌/超氧阴离子/一氧化氮Key words
Pelteobagrus fulvidraco/mannose receptor/Edwardsiella ictaluri/superoxide anion (O2-)/nitric oxide分类
生物科学引用本文复制引用
刘小玲,王虹,樊启学,兰江风,林蠡..黄颡鱼甘露糖受体基因的克隆和功能分析[J].水产学报,2017,41(7):1036-1043,8.基金项目
国家自然科学基金(31372563,31572657,31502199) （31372563,31572657,31502199）
中央高校基本科研业务费专项(2014PY035,2662015QC019) （2014PY035,2662015QC019）
湖北省科技支撑计划(2015BBA228,YSF2015000255) （2015BBA228,YSF2015000255）
农业部渔用药物创制重点实验室开放课题(201401) （201401）
广东省海洋与渔业局基金(A201512C003, 2015-115) （A201512C003, 2015-115）
武汉市科技局基金(2016020101010089)National Natural Science Foundation of China (31372563, 31572657, 31502199) （2016020101010089）
The Fundamental Research Funds for the Central Universities (2014PY035, 2662015QC019) （2014PY035, 2662015QC019）
Special Fund for Science and Technology from Hubei Province (2015BBA228,YSF2015000255) （2015BBA228,YSF2015000255）
Fund from Key Laboratory of Fishery Drug Development, Ministry of Agriculture (201401) （201401）
Special Funds from the Administration of Ocean and Fisheries of Guangdong Province (A201512C003,2015-115) （A201512C003,2015-115）
Fund from Wuhan Science and Technology Bureau (2016020101010089) （2016020101010089）
