The Fishery Institute of the Sichuan Academy of Agricultural Sciences, Chengdu, 611730, China.
The Fishery Institute of the Sichuan Academy of Agricultural Sciences, Chengdu, 611730, China.
Fish Shellfish Immunol. 2019 May;88:207-216. doi: 10.1016/j.fsi.2019.02.050. Epub 2019 Feb 23.
Antimicrobial peptides (AMPs) play essential roles in the innate immune system to protect against a wide variety of pathogens in aquatic environments. In this study, three very important AMPs, cathelicidin, hepcidin and defensin, were identified in the critically endangered Acipenser dabryanus. The full-length cDNA sequences of these three AMPs were identified from transcriptome sequencing and the rapid amplification of cDNA ends (RACE) technique. Phylogenetic analysis showed that cathelicidin formed a clade with the other members of the cathelicidin family, and similar results were obtained for hepcidin. The A. dabryanus β-defensin belonged to the fish class 2 β-defensins. A tissue distribution study showed that the three AMP transcripts could be detected constitutively in various tissues. The highest expression levels of cathelicidin and hepcidin were found in the liver, while defensin was primarily expressed in the skin. Bacterial challenge in vivo revealed significant changes in the gene expression of the three AMPs at both mucosal sites and systemic sites. Striking upregulation of cathelicidin and hepcidin was observed in the skin at 12 h post-challenge, with increases of more than 7000-fold and 1000-fold, respectively, compared to the control, and the expression of defensin mRNA was remarkably elevated in the hindgut (by 230-fold at 6 h post-challenge). Moreover, according to the expression profiles of the AMPs post-challenge, we found that the mucosal immune response occurred earlier than the systemic immune response following bacterial infection. Our results suggest that these three novel AMPs may play important roles in the innate immune system of A. dabryanus to protect against invading pathogens, especially during the mucosal immune response.
抗菌肽 (AMPs) 在先天免疫系统中发挥着重要作用,可抵御水生环境中的各种病原体。在这项研究中,从濒危物种达氏鲟的转录组测序和快速扩增 cDNA 末端 (RACE) 技术中鉴定出三种非常重要的 AMPs,即抗菌肽、hepcidin 和防御素。这三种 AMP 的全长 cDNA 序列从转录组测序和快速扩增 cDNA 末端 (RACE) 技术中鉴定出来。系统发育分析表明,抗菌肽与抗菌肽家族的其他成员形成一个分支,hepcidin 也得到了类似的结果。达氏鲟β-防御素属于鱼类 2 类β-防御素。组织分布研究表明,这三种 AMP 转录本可以在各种组织中持续检测到。抗菌肽和 hepcidin 的表达水平最高的组织是肝脏,而防御素主要在皮肤中表达。体内细菌攻毒实验表明,在黏膜和系统部位,三种 AMP 基因的表达均发生显著变化。皮肤中抗菌肽和 hepcidin 的基因表达在攻毒后 12 小时显著上调,与对照组相比,分别上调了 7000 倍和 1000 倍,防御素 mRNA 的表达在攻毒后 6 小时显著上调(增加了 230 倍)。此外,根据 AMP 攻毒后的表达谱,我们发现黏膜免疫反应比系统免疫反应更早发生在细菌感染后。我们的研究结果表明,这三种新型 AMP 可能在达氏鲟先天免疫系统中发挥重要作用,以抵御入侵病原体,尤其是在黏膜免疫反应中。