鲫鱼鳃对嗜水气单胞菌反应的综合转录组学和蛋白质组学分析

Xiucai Hu , Jie Bai , Rongrong Liu , Aijun Lv
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引用次数: 5

摘要

作为粘膜屏障之一,鱼鳃是抵御病原体感染的第一道防线。然而,鱼类对细菌感染的鳃粘膜免疫反应的确切机制仍需进一步研究。为了研究嗜水气单胞菌攻击鲫鱼(Carassius auratus)鳃粘膜免疫反应的病理变化和分子机制,利用RNA-seq的多组学分析和iTRAQ技术进行了转录组学和蛋白质组学研究。结果表明,鳃免疫反应主要与补体和凝血级联反应的激活、抗原处理和呈递、吞噬体、NOD样受体(NLR)和核因子κB(NFκB)信号通路有关。选择21种免疫相关DEG(即Clam、nfyal、snrpf、acin1b、psme、sf3b5、rbm8a、rbm25、prpf18、g3bp2、snrpd3l、tecrem-2、cfl-A、C7、lysC、ddx5、hsp90、α-2M、C9、C3和slc4a1a),通过qRT-PCR法验证其在嗜水气单胞菌感染中的免疫作用。同时,一些补体(C3、C7、C9、CFD、DF和FH)和抗原呈递(HSP90、MHCⅡ、CALR、CANX和PSME)蛋白显著参与了鳃组织对感染的防御过程,蛋白质-蛋白质相互作用(PPI)网络显示了这些DEP之间的免疫信号通路和相互作用。相关分析表明,iTRAQ和qRT-PCR结果显著相关(Pearson相关系数=0.70,p<;0.01)。据我们所知,首次通过多组学分析鉴定的鳃的转录组学和蛋白质组学有助于了解塞浦路斯物种局部粘膜免疫的分子机制。
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Comprehensive transcriptomics and proteomics analysis of Carassius auratus gills in response to Aeromonas hydrophila

As one of the mucosal barriers, fish gills represent the first line of defense against pathogen infection. However, the exact mechanism of gill mucosal immune response to bacterial infection still needs further investigation in fish. Here, to investigate pathological changes and molecular mechanisms of the mucosal immune response in the gills of crucian carp (Carassius auratus) challenged by Aeromonas hydrophila, the transcriptomics and proteomics were performed by using multi-omics analyses of RNA-seq coupled with iTRAQ techniques. The results demonstrated gill immune response were mostly related to the activation of complement and coagulation cascades, antigen processing and presentation, phagosome, NOD-like receptor (NLR) and nuclear factor κB (NFκB) signaling pathway. Selected 21 immune-related DEGs (ie., Clam, nfyal, snrpf, acin1b, psme, sf3b5, rbm8a, rbm25, prpf18, g3bp2, snrpd3l, tecrem-2, cfl-A, C7, lysC, ddx5, hsp90, α-2M, C9, C3 and slc4a1a) were verified for their immune roles in the A. hydrophila infection via using qRT-PCR assay. Meanwhile, some complement (C3, C7, C9, CFD, DF and FH) and antigen presenting (HSP90, MHC Ⅱ, CALR, CANX and PSME) proteins were significantly participated in the process of defense against infections in gill tissues, and protein-protein interaction (PPI) network displayed the immune signaling pathways and interactions among these DEPs. The correlation analysis indicated that the iTRAQ and qRT-PCR results was significantly correlated (Pearson's correlation coefficient = 0.70, p < 0.01). To our knowledge, the transcriptomics and proteomics of gills firstly identified by multi-omics analyses contribute to understanding on the molecular mechanisms of local mucosal immunity in cyprinid species.

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