Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture, Shanghai, 201306, China; Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding Genetics, Shanghai, 201306, China; Laboratory of Cell and Molecular Biology, Shanghai Ocean University, Shanghai, 201306, China; Key Laboratory of Aquatic Genomics, Ministry of Agriculture, Beijing, 100141, China.
Key Laboratory of Freshwater Aquatic Genetic Resources, Ministry of Agriculture, Shanghai, 201306, China; Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding Genetics, Shanghai, 201306, China; Laboratory of Cell and Molecular Biology, Shanghai Ocean University, Shanghai, 201306, China.
Fish Shellfish Immunol. 2019 Oct;93:183-190. doi: 10.1016/j.fsi.2019.07.055. Epub 2019 Jul 19.
In mammals, a matricellular protein, thrombospondin 2 (Thbs2) has been reported to play important roles in modulating cell-matrix interactions, vascular integrity and thrombosis formation. However, the role of gene, thbs2 has not yet been studied in teleost. In the present study, this novel fish gene from Japanese flounder was cloned and its function in resistant to lymphocystis disease virus was elucidated. The Japanese flounder thbs2 encoded a 1176-amino acid protein with 91% identity to medaka. Amino acid sequence indicated that Japanese flounder Thbs2 contained 10 typical conserved domains. The thbs2 was expressed in all stages of embryo development, and in hatched larva stage, its expression was significantly higher than that in other stages (P < 0.05). The relative expression level of thbs2 was significantly higher in the head kidney, liver, blood, gill, and heart of the lymphocystis disease virus resistant fish than in sensitive fish (P < 0.05); and in muscle, this difference was at highly significant (P < 0.01). Additionally, the distribution of Thbs2 in tissue was evaluated by immunohistochemical staining. Subcellular localization analysis showed that Thbs2 was distributed throughout the cytoplasm of the cells. Taken together, our results provide new basic data for thbs2 function, especially its role in anti-lymphocystis disease virus immune response.
在哺乳动物中,一种细胞外基质蛋白,血小板反应蛋白 2(Thbs2)被报道在调节细胞-基质相互作用、血管完整性和血栓形成中发挥重要作用。然而,该基因在硬骨鱼中的作用尚未研究。在本研究中,从牙鲆克隆了这种新型鱼类基因,并阐明了其在抵抗淋巴囊肿病病毒中的功能。牙鲆 thbs2 编码一个 1176 个氨基酸的蛋白质,与青鳉的同源性为 91%。氨基酸序列表明,牙鲆 Thbs2 包含 10 个典型的保守结构域。thbs2 在胚胎发育的所有阶段都有表达,在孵化的幼鱼阶段,其表达水平显著高于其他阶段(P<0.05)。在抗淋巴囊肿病病毒鱼的头肾、肝脏、血液、鳃和心脏中,thbs2 的相对表达水平显著高于敏感鱼(P<0.05);在肌肉中,这种差异达到极显著水平(P<0.01)。此外,通过免疫组织化学染色评估了 Thbs2 在组织中的分布。亚细胞定位分析表明,Thbs2 分布在细胞的整个细胞质中。总之,我们的结果为 thbs2 的功能提供了新的基础数据,特别是其在抗淋巴囊肿病病毒免疫反应中的作用。