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基于iTRAQ的感染病毒1的加勒比刺龙虾肝胰腺蛋白质组学分析:代谢和生理意义

iTRAQ-Based Proteomic Profile Analysis of the Hepatopancreas of Caribbean Spiny Lobsters Infected With Virus 1: Metabolic and Physiological Implications.

作者信息

Zamora-Briseño Jesús Alejandro, Ruiz-May Eliel, Elizalde-Contreras José Miguel, Hernández-Velázquez Ioreni Margarita, Hernández-Pérez Ariadne, Fuentes-García Ana Guadalupe, Herrera-Salvatierra Nancy, Briones-Fourzán Patricia, Pascual-Jiménez Cristina, Lozano-Álvarez Enrique, Rodríguez-Canul Rossanna

机构信息

Laboratorio de Inmunología y Biología Molecular, Departamento de Recursos del Mar, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional-Unidad Mérida, Mérida, Mexico.

Instituto de Ecología, Red de Estudios Moleculares Avanzados, Clúster Científico y Tecnológico BioMimic®, Xalapa, Mexico.

出版信息

Front Microbiol. 2020 May 29;11:1084. doi: 10.3389/fmicb.2020.01084. eCollection 2020.

Abstract

The Caribbean spiny lobster (Latreille, 1084) sustains economically valuable fisheries throughout the wider Caribbean region. This species is currently affected by the pathogenic virus Virus 1 (PaV1) that causes a systemic and chronic-degenerative infection in juvenile spiny lobsters . To date, there is no available information regarding the host alterations induced by this pathogen at the molecular level. In the present study, comparative proteomic analyses of the changes in the hepatopancreas between infected and non-infected juvenile lobsters were analyzed by isobaric tags for relative and absolute quantitation (iTRAQ) coupled to synchronous precursor selection (SPS)-based MS. We identified a total of 636 proteins, being 68 down-regulated and 71 up-regulated proteins. Among the down-regulated proteins, we identified several enzymes involved in the metabolism of hormones and lipids, digestive proteases and glycosidases, while proteins associated with the histone core, protein synthesis, immune response and RNA regulation were up-regulated. Several misregulated enzymes involved in the regulation of neuromodulators were also identified. RT-qPCR assays were used to validate the expression of transcripts encoding for selected differential proteins that were in concordance to proteomic data, as well as the tendency observed in the enzymatic activities of trypsin, chymotrypsin, and glycosidase. In a similar way, we observed glycogen reduction in muscle, and an increase in plasma acylglycerides and glucose, which may be explained by proteomic data. This study provides the first insight into the molecular changes in the hepatopancreas of Caribbean spiny lobsters associated to PaV1 infection. Data provided herein would help to clarify the origin of the molecular misregulations observed at macroscopic level in this host-pathogen interaction.

摘要

加勒比刺龙虾(Latreille,1804年)支撑着整个加勒比海广大区域具有经济价值的渔业。该物种目前受到致病性病毒——病毒1(PaV1)的影响,这种病毒会在幼年刺龙虾中引发全身性和慢性退行性感染。迄今为止,尚无关于这种病原体在分子水平上引起的宿主变化的可用信息。在本研究中,通过等压标签相对和绝对定量(iTRAQ)结合基于同步前体选择(SPS)的质谱分析,对受感染和未受感染的幼年龙虾肝胰腺变化进行了比较蛋白质组学分析。我们总共鉴定出636种蛋白质,其中68种蛋白质下调,71种蛋白质上调。在下调的蛋白质中,我们鉴定出了几种参与激素和脂质代谢、消化蛋白酶和糖苷酶的酶,而与组蛋白核心、蛋白质合成、免疫反应和RNA调节相关的蛋白质则上调。还鉴定出了几种参与神经调节剂调节的失调酶。实时定量聚合酶链反应(RT-qPCR)分析用于验证所选差异蛋白质编码转录本的表达,这些转录本与蛋白质组学数据一致,同时也验证了胰蛋白酶、胰凝乳蛋白酶和糖苷酶的酶活性变化趋势。同样,我们观察到肌肉中糖原减少,血浆甘油酯和葡萄糖增加,这可以用蛋白质组学数据来解释。本研究首次深入了解了与PaV1感染相关的加勒比刺龙虾肝胰腺的分子变化。本文提供的数据将有助于阐明在这种宿主-病原体相互作用中在宏观水平上观察到的分子失调的起源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bf76/7273172/3903b179970c/fmicb-11-01084-g001.jpg

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