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对副溶血弧菌引起的凡纳滨对虾急性肝胰腺坏死病的转录组分析。

Transcriptomic analysis of Pacific white shrimp (Litopenaeus vannamei, Boone 1931) in response to acute hepatopancreatic necrosis disease caused by Vibrio parahaemolyticus.

机构信息

Laboratorio de Patogénesis Microbiana, Centro de Investigaciones Biológicas del Noroeste, S. C. (CIBNOR), La Paz, Baja California Sur, México.

Programa de Cátedras CONACyT, Centro de Investigaciones Biológicas del Noroeste, S. C. (CIBNOR), La Paz, Baja California Sur, México.

出版信息

PLoS One. 2019 Aug 13;14(8):e0220993. doi: 10.1371/journal.pone.0220993. eCollection 2019.

Abstract

Acute hepatopancreatic necrosis disease (AHPND), caused by marine bacteria Vibrio Parahaemolyticus, is a huge problem in shrimp farms. The V. parahaemolyticus infecting material is contained in a plasmid which encodes for the lethal toxins PirABVp, whose primary target tissue is the hepatopancreas, causing sloughing of epithelial cells, necrosis, and massive hemocyte infiltration. To get a better understanding of the hepatopancreas response during AHPND, juvenile shrimp Litopenaeus vannamei were infected by immersion with V. parahaemolyticus. We performed transcriptomic mRNA sequencing of infected shrimp hepatopancreas, at 24 hours post-infection, to identify novel differentially expressed genes a total of 174,098 transcripts were examined of which 915 transcripts were found differentially expressed after comparative transcriptomic analysis: 442 up-regulated and 473 down-regulated transcripts. Gene Ontology term enrichment analysis for up-regulated transcripts includes metabolic process, regulation of programmed cell death, carbohydrate metabolic process, and biological adhesion, whereas for down-regulated transcripts include, microtubule-based process, cell activation, and chitin metabolic process. The analysis of protein- protein network between up and down-regulated genes indicates that the first gene interactions are connected to oxidation-processes and sarcomere organization. Additionally, protein-protein networks analysis identified 20-top highly connected hub nodes. Based on their immunological or metabolic function, ten candidate transcripts were selected to measure their mRNA relative expression levels in AHPND infected shrimp hepatopancreas by RT-qPCR. Our results indicate a close connection between the immune and metabolism systems during AHPND infection. Our RNA-Seq and RT-qPCR data provide the possible immunological and physiological scenario as well as the molecular pathways that take place in the shrimp hepatopancreas in response to an infectious disease.

摘要

急性肝胰腺坏死病(AHPND)由海洋细菌副溶血弧菌引起,是对虾养殖场的一个巨大问题。感染的副溶血弧菌材料包含在一个质粒中,该质粒编码致死毒素 PirABVp,其主要靶组织是肝胰腺,导致上皮细胞脱落、坏死和大量血细胞浸润。为了更好地了解 AHPND 期间肝胰腺的反应,我们通过浸泡感染了凡纳滨对虾(Litopenaeus vannamei)幼虾。我们对感染虾肝胰腺的转录组 mRNA 进行了测序,在感染后 24 小时,总共检查了 174098 个转录本,通过比较转录组分析发现了 915 个差异表达基因:442 个上调和 473 个下调转录本。上调转录本的基因本体论术语富集分析包括代谢过程、程序性细胞死亡的调节、碳水化合物代谢过程和生物粘附,而下调转录本包括微管过程、细胞激活和几丁质代谢过程。上调和下调基因之间的蛋白质-蛋白质网络分析表明,第一个基因相互作用与氧化过程和肌节组织有关。此外,蛋白质-蛋白质网络分析确定了 20 个高度连接的枢纽节点。基于它们的免疫或代谢功能,选择了 10 个候选转录本,通过 RT-qPCR 测量它们在 AHPND 感染虾肝胰腺中的 mRNA 相对表达水平。我们的结果表明,在 AHPND 感染期间,免疫和代谢系统之间存在密切联系。我们的 RNA-Seq 和 RT-qPCR 数据提供了可能的免疫和生理情况以及分子途径,这些途径在虾肝胰腺中对传染病发生反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51e1/6692014/b0d28d1314b1/pone.0220993.g001.jpg

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