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环孢素 A 基因在黄颡鱼对爱德华氏菌抗性中的功能与关联分析。

Function and association analysis of Cyclophilin A gene with resistance to Edwardsiella ictaluri in yellow catfish.

机构信息

College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, China.

Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072, China.

出版信息

Dev Comp Immunol. 2020 Dec;113:103783. doi: 10.1016/j.dci.2020.103783. Epub 2020 Jul 29.

Abstract

Edwardsiella ictaluri (E. ictaluri) is one of the main bacterial pathogens in catfish which has caused serious economic loss to yellow catfish (Pelteobagrus fulvidraco) in China. In our previous work, we demonstrated that CypA was up-regulated at the early stage of E. ictaluri infection in yellow catfish and displayed strong chemotactic activity for leukocytes in vitro. However, the effect of CypA on E. ictaluri is unknown in vivo. Therefore, two homozygous transgenic zebrafish lines expressing yellow catfish CypA (TG-CypA-1 and TG-CypA-2) were generated. After challenged with E. ictaluri at a dose of 1.0 × 10 CFU per adult fish, both two transgenic lines exhibited a higher resistance to bacterial infection than the wildtype zebrafish. Herein, CypA gene in E. ictaluri-challenged yellow catfish was screened for presence of polymorphisms by sequencing and six single nucleotide polymorphisms (SNPs) were identified. SNP association analysis revealed that 528T/C SNP in the first intron was significantly different in disease-susceptible and -resistant groups, which was confirmed in two independent populations of yellow catfish. Moreover, the relative expression of CypA in the resistant group (CC genotype in 528T/C SNP) was significantly higher than that in the susceptible group (TT genotype in 528T/C SNP) in different immune organs of yellow catfish including spleen, head kidney, body kidney and liver. Our results reveal the potential function of CypA in host defense to bacterial infection and suggest the SNP marker in CypA gene associated with the resistance to E. ictaluri may facilitate the selective breeding of disease-resistant yellow catfish in the future.

摘要

爱德华氏菌(E. ictaluri)是一种鱼类病原菌,是导致中国黄颡鱼(Pelteobagrus fulvidraco)严重经济损失的主要细菌病原体之一。在我们之前的工作中,我们证明 CypA 在黄颡鱼感染爱德华氏菌的早期阶段上调,并在体外显示出对白细胞强烈的趋化活性。然而,CypA 在体内对爱德华氏菌的影响尚不清楚。因此,我们生成了两条表达黄颡鱼 CypA 的纯合转基因斑马鱼系(TG-CypA-1 和 TG-CypA-2)。在以每尾成鱼 1.0×10 CFU 的剂量受到爱德华氏菌攻击后,这两条转基因系均表现出比野生型斑马鱼更高的抗细菌感染能力。在此,通过测序筛选出爱德华氏菌感染黄颡鱼 CypA 基因中的多态性,共鉴定出 6 个单核苷酸多态性(SNP)。SNP 关联分析显示,第一内含子中的 528T/C SNP 在易感和抗性组之间存在显著差异,在黄颡鱼的两个独立群体中得到了验证。此外,在黄颡鱼的不同免疫器官中,包括脾脏、头肾、体肾和肝脏,528T/C SNP 中的抗性组(CC 基因型)的 CypA 相对表达量明显高于易感组(TT 基因型)。我们的研究结果揭示了 CypA 在宿主防御细菌感染中的潜在功能,并提示 CypA 基因中的 SNP 标记与对爱德华氏菌的抗性相关,这可能有助于未来对黄颡鱼进行抗病性的选择性繁殖。

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