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杂交黄颡鱼(黄颡鱼♀×瓦氏黄颡鱼♂)中fadd基因对外源嗜水气单胞菌或迟缓爱德华氏菌刺激的特征分析及表达谱研究

Characterization and expression profiling of fadd gene in response to exogenous Aeromonas hydrophila or Edwardsiella tarda challenge in the hybrid yellow catfish (Pelteobagrus fulvidraco ♀ × P. vachelli ♂).

作者信息

Guo Shengtao, Zeng Mengsha, Chen Lei, Chen Hanxi, Gao Wenxue, Wang Zhongyi, Shi Qiong, Song Zhaobin

机构信息

Key Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610065, China.

Shenzhen Key Lab of Marine Genomics, Guangdong Provincial Key Lab of Molecular Breeding in Marine Economic Animals, BGI Academy of Marine Sciences, BGI Marine, BGI, Shenzhen 518083, China.

出版信息

Fish Shellfish Immunol. 2023 Oct;141:109021. doi: 10.1016/j.fsi.2023.109021. Epub 2023 Aug 24.

Abstract

In mammals, fas-associated protein with death domain (FADD) is involved in the process of cell apoptosis and plays a key role in innate immune signaling. Nevertheless, its detailed molecular mechanisms underlying apoptosis and immune responses to exogenous bacterial infections in teleosts remain largely unknown. In this study, a group of 60 hybrid yellow catfish (with the body weight of 25 ± 0.5 g) were used in subsequent experiments, we examined the expression profiling of fadd gene through comparative genomics and comparative immunological methods. Our results showed that fadd in the hybrid yellow catfish (hycfadd) exhibited similar gene and spatial structures to those in other vertebrates, and formed an independent clade in phylogeny. An expression pattern analysis revealed that hycfadd widely transcribed in various tissues, with the highest transcription level in the liver. Furthermore, expression profiling of hycfadd when intraperitoneally infected with 50 μL of exogenous Aeromonas hydrophila (2.0 × 10 CFU/mL) or Edwardsiella tarda (2.0 × 10 CFU/mL) within 48 h were significantly up-regulated in the kidney, spleen, liver and intestine. Important genes in the toll like receptor (tlr) 1-tlr2- myeloid differentiation primary response 88 (MyD88)-fadd-caspase (casp) 8 cascades of TLR signaling pathway in liver were significantly up-regulated after the A. hydrophila stimulation, suggesting that apoptosis through the TLR signaling pathway may have been triggered and activated, which were further verified in the liver, kidney, spleen, intestine and gill by a TUNEL assay. Overall, this study provides solid evidence for the bacterial induction of fadd-related apoptosis in teleosts.

摘要

在哺乳动物中,死亡结构域相关蛋白(FADD)参与细胞凋亡过程,并在先天免疫信号传导中起关键作用。然而,硬骨鱼对外源细菌感染的凋亡及免疫反应的详细分子机制仍不清楚。本研究选用60尾杂交黄颡鱼(体重25±0.5 g)进行后续实验,通过比较基因组学和比较免疫学方法检测fadd基因的表达谱。结果显示,杂交黄颡鱼fadd(hycfadd)与其他脊椎动物的fadd具有相似的基因和空间结构,且在系统发育中形成一个独立分支。表达模式分析表明,hycfadd在各组织中广泛转录,在肝脏中转录水平最高。此外,在48小时内腹腔注射50 μL外源嗜水气单胞菌(2.0×10 CFU/mL)或迟缓爱德华氏菌(2.0×10 CFU/mL)后,hycfadd在肾脏、脾脏、肝脏和肠道中的表达谱显著上调。嗜水气单胞菌刺激后,肝脏中Toll样受体(TLR)1-TLR2-髓样分化初级反应88(MyD88)-fadd-半胱天冬酶(casp)8级联反应中的重要基因显著上调,提示可能通过TLR信号通路触发并激活了凋亡,TUNEL检测在肝脏、肾脏、脾脏、肠道和鳃中进一步验证了这一点。总体而言,本研究为硬骨鱼中细菌诱导的fadd相关凋亡提供了确凿证据。

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