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锯缘青蟹成纤维细胞生长因子受体(FGFR)的鉴定与功能分析:FGFR 参与甲壳动物固有免疫的证据。

Identification and function analysis of two fibroblast growth factor receptor (FGFR) from Scylla paramamosain: The evidence of FGFR involved in innate immunity in crustacean.

机构信息

College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.

College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China.

出版信息

Fish Shellfish Immunol. 2022 Dec;131:602-611. doi: 10.1016/j.fsi.2022.08.075. Epub 2022 Sep 3.

DOI:10.1016/j.fsi.2022.08.075
PMID:36064005
Abstract

The fibroblast growth factor receptor (FGFR) belongs to the tyrosine kinase family consisting of four members (FGFR1-4). This study involved identification and characterization of FGFR1 and FGFR3 from mud crab Scylla paramamosain for the first time. The obtained cDNAs of SpFGFR1 and SpFGFR3 were 2,380 bp and 2,982 bp in length with a 1,503 bp and 2,310 bp open reading frame, respectively. The predicted SpFGFR1 protein included three immunoglobulin domains and a transmembrane region, while SpFGFR3 protein possessed a typical TyrKc (Tyrosine kinase, catalytic) domain. Real-time PCR analysis showed that SpFGFR1 and SpFGFR3 were highly expressed in the hepatopancreas. Furthermore, the expression levels of SpFGFR1 and SpFGFR3 in the hepatopancreas were enhanced following challenges with Vibro alginolyticus, Staphylococcus aureus, Poly (I:C) and White spot syndrome virus, which shows the involvement of SpFGFR1 and SpFGFR3 in innate immune response to infections from bacteria and virus. There was significant suppression of six antimicrobial peptide genes (SpALF1-5 and SpCrustin) and three NF-κB members (SpDorsal, SpIKK and SpRelish) when SpFGFR1 and SpFGFR3 was interfered in vivo. Also, treatment of the hemocytes with specific inhibitor of SpFGFR for 24 h consistently down-regulated SpDorsal, SpRelish and AMPs. These results suggested that SpFGFR1 and SpFGFR3 played important roles in regulating the Toll signaling pathway and immune deficiency (IMD) pathway through NF-κB signaling pathway. These findings may provide new insights into the role of FGFRs in the innate immune function of crustaceans.

摘要

成纤维细胞生长因子受体(FGFR)属于酪氨酸激酶家族,由四个成员(FGFR1-4)组成。本研究首次从泥蟹锯缘青蟹中鉴定和表征了 FGFR1 和 FGFR3。获得的 SpFGFR1 和 SpFGFR3 cDNA 分别长 2380bp 和 2982bp,开放阅读框分别为 1503bp 和 2310bp。预测的 SpFGFR1 蛋白包含三个免疫球蛋白结构域和一个跨膜区,而 SpFGFR3 蛋白具有典型的 TyrKc(酪氨酸激酶,催化)结构域。实时 PCR 分析显示,SpFGFR1 和 SpFGFR3 在肝胰腺中高度表达。此外,肝胰腺中 SpFGFR1 和 SpFGFR3 的表达水平在受到弧菌、金黄色葡萄球菌、聚肌胞和白斑综合征病毒的挑战后增强,表明 SpFGFR1 和 SpFGFR3 参与了对细菌和病毒感染的先天免疫反应。当体内干扰 SpFGFR1 和 SpFGFR3 时,六个抗菌肽基因(SpALF1-5 和 SpCrustin)和三个 NF-κB 成员(SpDorsal、SpIKK 和 SpRelish)的表达显著受到抑制。此外,用 SpFGFR 的特异性抑制剂处理 24 小时的血细胞一致地下调了 SpDorsal、SpRelish 和 AMPs。这些结果表明,SpFGFR1 和 SpFGFR3 通过 NF-κB 信号通路在调节 Toll 信号通路和免疫缺陷(IMD)通路中发挥重要作用。这些发现可能为 FGFRs 在甲壳动物先天免疫功能中的作用提供新的见解。

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