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黄颡鱼(Pelteobagrus fulvidraco) crtc2 基因的分子特征、时空表达模式及其免疫作用。

Molecular characterization, spatio-temporal expression patterns of crtc2 gene and its immune roles in yellow catfish (Pelteobagrus fulvidraco).

机构信息

Key Laboratory of Sichuan Province for Fishes Conservation and Utilization in the Upper Reaches of the Yangtze River, Neijiang Normal University, Neijiang, 641100, China; College of Life Science, Neijiang Normal University, Neijiang, 641100, China; School of Animal Science, Yangtze University, Jingzhou, 424020, China.

Key Laboratory of Sichuan Province for Fishes Conservation and Utilization in the Upper Reaches of the Yangtze River, Neijiang Normal University, Neijiang, 641100, China; College of Life Science, Neijiang Normal University, Neijiang, 641100, China; School of Animal Science, Yangtze University, Jingzhou, 424020, 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. 2024 Nov;154:109877. doi: 10.1016/j.fsi.2024.109877. Epub 2024 Sep 7.

Abstract

cAMP response element binding (CREB) protein 2 (CRTC2) is a transcriptional coactivator of CREB and plays an important role in the immune system. Thus far, the physiological roles of Crtc2 in teleost are still poorly understood. In this study, the crtc2 gene was identified and characterized from yellow catfish (Pelteobagrus fulvidraco; therefore, the gene is termed as pfcrtc2), and its evolutionary and molecular characteristics as well as potential immunity-related roles were investigated. Our results showed that the open reading frame of pfcrtc2 was 2346 bp in length, encoding a protein with 781 amino acids. Gene structure analysis revealed its existence of 14 exons and 13 introns. A phylogenetic analysis proved that the tree of crtc2 was clustered into five groups, exhibiting a similar evolutionary topology with species evolution. Multiple protein sequences alignment demonstrated high conservation of the crtc2 in various vertebrates with similar structure. Syntenic and gene structural comparisons further established that crtc2 was highly conserved, implying its similar roles in diverse vertebrates. Tissue distribution pattern detected by quantitative real-time PCR showed that the pfcrtc2 gene was almost expressed in all detected tissues except for eyes, with the highest expression levels in the gonad, indicating that Crtc2 may play important roles in various tissues. In addition, pfcrtc2 was transcribed at all developmental stages in yellow catfish, showing the highest expression levels at 12 h after fertilization. Finally, the transcriptional profiles of crtc2 were significantly increased in yellow catfishes injected with Aeromonas hydrophila or Poly I:C, which shared a consistent change pattern with four immune-related genes including IL-17A, IL-10, MAPKp38, and NF-κBp65, suggesting pfCrtc2 may play critical roles in preventing both exogenous bacteria and virus invasion. In summary, our findings lay a solid foundation for further studies on the functions of pfcrtc2, and provide novel genetic loci for developing new strategies to control disease outbreak in teleost.

摘要

环磷酸腺苷反应元件结合蛋白 2(CREB)是 CREB 的转录共激活因子,在免疫系统中发挥着重要作用。迄今为止,硬骨鱼类 Crtc2 的生理功能仍知之甚少。本研究从黄颡鱼(Pelteobagrus fulvidraco)中鉴定并表征了 crtc2 基因(因此将该基因命名为 pfcrtc2),并研究了其进化和分子特征及其潜在的免疫相关作用。我们的结果表明,pfcrtc2 的开放阅读框长 2346bp,编码一个含有 781 个氨基酸的蛋白质。基因结构分析表明其存在 14 个外显子和 13 个内含子。系统进化分析表明,crtc2 树聚为五个分支,与物种进化具有相似的进化拓扑结构。多种蛋白质序列比对表明,crtc2 在各种脊椎动物中具有高度的保守性,具有相似的结构。基因结构和基因共线性比较进一步表明,crtc2 在不同的脊椎动物中高度保守,暗示其在不同脊椎动物中具有相似的作用。实时定量 PCR 检测的组织分布模式表明,pfcrtc2 基因几乎在所有检测组织中均有表达,除眼睛外,性腺中的表达水平最高,表明 Crtc2 可能在各种组织中发挥重要作用。此外,pfcrtc2 在黄颡鱼的所有发育阶段均有转录,在受精后 12h 时表达水平最高。最后,Aeromonas hydrophila 或 Poly I:C 注射后,crtc2 的转录谱显著增加,与四个免疫相关基因(包括 IL-17A、IL-10、MAPKp38 和 NF-κBp65)的变化趋势一致,表明 pfCrtc2 可能在预防外源性细菌和病毒入侵中发挥关键作用。综上所述,本研究为进一步研究 pfcrtc2 的功能奠定了基础,为开发控制硬骨鱼类疾病爆发的新策略提供了新的遗传位点。

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