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斑节对虾β整合素的全面表达分析表明其参与先天免疫和氨氮胁迫反应。

Comprehensive expression analysis of the beta integrin from Penaeus monodon indicating its participation in innate immunity and ammonia nitrogen stress response.

机构信息

South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture, Guangzhou, 510300, PR China; College of Aqua-life Science and Technology, Shanghai Ocean University, Shanghai, PR China.

South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Key Laboratory of South China Sea Fishery Resources Exploitation and Utilization, Ministry of Agriculture, Guangzhou, 510300, PR China; Shenzhen Base of South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shenzhen, 518108, PR China.

出版信息

Fish Shellfish Immunol. 2020 Mar;98:887-898. doi: 10.1016/j.fsi.2019.11.049. Epub 2019 Nov 23.

Abstract

The aim of the present study was to investigate the function of the beta integrin (PmItgb) in Penaeus monodon. The 3011 bp cDNA sequence of PmItgb was cloned from P. monodon using rapid amplification of cDNA ends (RACE) PCR. Phylogenetic tree analyses indicated that the amino acid sequence of PmItgb should be merged into Fenneropenaeus chinensis (93%). Quantitative real-time PCR (q RT-PCR) revealed that PmItgb mRNA was highly expressed in the hemocytes. In addition, with regard to developmental stages, PmItgb showed significantly higher expression in oosperm, nauplius IV, zoea I and III, and post larval stages than that in other development stages. PmItgb expression in the shrimp epidermis was higher in the postmolt (B) stage, and lower in other molting stages. We also found that Vibrio harveyi and V. anguillarum challenge enhanced PmItgb expression in the hepatopancreas and gills. When PmItgb was inhibited, innate immunity-related genes such as ALF, crustin 1, crustin 7, penaeidin 3, and penaeidin 5 were significantly down-regulated. Furthermore, we demonstrated that PmItgb knock-down by specific dsRNA reduced bacterial clearance. In high ammonia nitrogen concentrations, PmItgb was significantly up-regulated in the hepatopancreas and gills. After PmItgb was silenced, the rate of mortality owing to high ammonia nitrogen concentrations decreased; the expression of related anti-apoptotic genes was up-regulated, and that of the apoptotic genes was slightly down-regulated. These results suggested that PmItgb may be involved in shrimp innate immunity and mediate apoptosis of hepatopancreatic cells induced by high ammonia nitrogen environments.

摘要

本研究旨在探究β整合素(PmItgb)在斑节对虾中的功能。采用快速扩增 cDNA 末端(RACE)PCR 技术,从斑节对虾中克隆出 3011bp 的 PmItgb cDNA 序列。系统进化树分析表明,PmItgb 氨基酸序列应与中国明对虾(93%)融合。定量实时 PCR(q RT-PCR)显示,PmItgb mRNA 在血细胞中高度表达。此外,就发育阶段而言,PmItgb 在卵母细胞、无节幼体 IV 期、幼体 I 期和 III 期以及幼体后期的表达显著高于其他发育阶段。在虾的表皮中,PmItgb 在蜕皮后期(B 期)的表达较高,而在其他蜕皮阶段的表达较低。我们还发现,哈维弧菌和鳗弧菌的刺激增强了 PmItgb 在肝胰腺和鳃中的表达。当 PmItgb 受到抑制时,如 ALF、壳聚糖 1、壳聚糖 7、抗菌肽 3 和抗菌肽 5 等先天免疫相关基因的表达显著下调。此外,我们证明了通过特异性 dsRNA 抑制 PmItgb 会降低细菌清除率。在高氨氮浓度下,PmItgb 在肝胰腺和鳃中显著上调。沉默 PmItgb 后,由于高氨氮浓度导致的死亡率降低;相关抗凋亡基因的表达上调,凋亡基因的表达略有下调。这些结果表明,PmItgb 可能参与虾的先天免疫,并介导高氨氮环境诱导的肝胰腺细胞凋亡。

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