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SsC1qDC基因在毛蚶免疫防御中的作用
彭强1,2,3, 王进京2, 王春德2, 卢霞2, 刘相全3, 刘桂龙4, 徐鑫4, 许贺5,6, 宁军号2
1.上海海洋大学 水产与生命学院, 上海 201306;2.中国科学院烟台海岸带研究所, 山东 烟台 264003;3.山东海洋资源与环境研究所, 山东 烟台 264006;4.烟台海之春水产种业有限公司, 山东 烟台 264006;5.江苏宝源生物科技有限公司, 江苏 连云港 222144;6.江苏海泰海洋科技有限公司, 江苏 连云港 222144
摘要:
含C1q结构域蛋白(C1qDCs)在无脊椎动物免疫应答中发挥重要作用, 但目前有关毛蚶(Scapharca subcrenata)C1qDCs免疫功能的研究鲜有报道。本研究从毛蚶中鉴定出一个含有典型C1q结构域的基因(命名为SsC1qDC), 并分析了其对主要病原感染的响应特征。SsC1qDC的cDNA全长序列为711 bp, 编码含有N端信号肽和199个氨基酸的蛋白,系统进化分析显示SsC1qDC与双壳贝类的C1qDCs聚在一起。SsC1qDC在毛蚶卵细胞到眼点幼虫时期的表达量随发育阶段显著增加, 且该基因在成体毛蚶各组织中均有表达, 其中在肝胰腺和闭壳肌中表达水平较高。副溶血弧菌刺激可显著诱导SsC1qDC基因高表达, 敲降SsC1qDC基因表达后毛蚶死亡率和血细胞凋亡水平显著升高, 且导致毛蚶血细胞总数和吞噬活力显著下降。同时, 敲降SsC1qDC基因的表达可显著抑制母源免疫相关基因VgLSZDscamTEPSsC1qDC-3的表达, 而C3PAF3SsC1qDC-1SsC1qDC-2等基因的表达显著上调。本研究结果表明, SsC1qDC在毛蚶抗弧菌响应中发挥重要作用, 对贝类免疫学和抗病新品系的选育有重要的指导意义。
关键词:  SsC1qDC  毛蚶(Scapharca subcrenata)  表达图谱  RNA干扰  免疫防御
DOI:10.11759/hykx20230518001
分类号:
基金项目:国家自然科学基金资助项目(31972791); 山东省农业良种工程资助项目(2020LZGC016); 山东省烟台市科技项目专项基金资助项目(2022XCZX083); 山东省现代农业产业技术体系建设项目(SDAIT-14)
A novel C1q domain-containing gene (SsC1qDC) from Scapharca subcrenata participates in the immune defense against pathogen infection
PENG Qiang1,2,3, WANG Jin-jing2, WANG Chun-de2, LU Xia2, LIU Xiang-quan3, LIU Gui-long4, XU Xin4, XU He5,6, NING Jun-hao2
1.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;2.Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai 264003, China;3.Shandong Marine Resource and Environment Research Institute, Yantai 264006, China;4.Yantai Spring-Sea AquaSeed Co., Ltd., Yantai 264006, China;5.Jiangsu Baoyuan Biotechnology Co., Ltd., Lianyungang 222144, China;6.Jiangsu Haitai MariTech Co., Ltd., Lianyungang 222144, China
Abstract:
The C1q domain-containing proteins (C1qDCs) have been demonstrated to play crucial roles in the immune responses of invertebrates, with very few reports in the ark shells. In the present study, a C1qDC gene with a typical C1q domain (designated as SsC1qDC) was identified from the ark shell Scapharca subcrenata and its possible immune responses against pathogen infection was also characterized. The full-length cDNA sequence of SsC1qDC was 711 bp, encoding a peptide of 199 amino acids with an N-terminal signal peptide. SsC1qDC was clustered with the C1qDCs from bivalves in the phylogenetic tree. SsC1qDC was widely expressed in the tissues, with the high expression level in hepatopancreas and adductor muscles. The expression of SsC1qDC was increased significantly from eggs to eye-point larvae stage and induced remarkably by Vibro parahaemolyticus infection. Moreover, compared to the controls, knockdown of SsC1qDC resulted in a remarkable increase of mortality and apoptotic levels of ark shells after V. parahaemolyticus challenge, and thus a significant reduction of total number and phagocytosis of hemocytes. Meanwhile, knockdown of SsC1qDC could significantly inhibit the expression of maternally derived immune-related genes (Vg, LSZ, Dscam, TEP and SsC1qDC-3), while the expression of C3, PAF3, SsC1qDC-1 and SsC1qDC-2 was up-regulated remarkably. Taken together, these results suggested that SsC1qDC performs crucial roles in immune defense against pathogen infection in ark shells, which is of great guiding significance to the immunology of shellfish and the breeding of new disease-resistant strains.
Key words:  SsC1qDC  Scapharca subcrenata  expression profiles  RNA interference  immune defense
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