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登革热病毒2型感染会改变亚洲虎蚊(白纹伊蚊)中肠和胴体的基因表达。

Dengue virus serotype 2 infection alters midgut and carcass gene expression in the Asian tiger mosquito, Aedes albopictus.

作者信息

Tsujimoto Hitoshi, Hanley Kathryn A, Sundararajan Anitha, Devitt Nicholas P, Schilkey Faye D, Hansen Immo A

机构信息

Department of Biology, New Mexico State University, Las Cruces, New Mexico, United States of America.

NM-INBRE Sequencing and Bioinformatics Core, National Center for Genome Resources, Santa Fe, New Mexico, United States of America.

出版信息

PLoS One. 2017 Feb 2;12(2):e0171345. doi: 10.1371/journal.pone.0171345. eCollection 2017.

Abstract

BACKGROUND

The Asian tiger mosquito, Aedes albopictus is currently an important vector for dengue, chikungunya and Zika virus, and its role in transmission of arthropod-borne viruses (arboviruses) may increase in the future due to its ability to colonize temperate regions. In contrast to Aedes aegypti, the dominant vector of dengue, chikungunya and Zika virus, genetic responses of Ae. albopictus upon infection with an arbovirus are not well characterized. Here we present a study of the changes in transcript expression in Ae. albopictus exposed to dengue virus serotype 2 via feeding on an artificial bloodmeal.

METHODOLOGY/PRINCIPAL FINDINGS: We isolated midguts and midgut-free carcasses of Ae. albopictus fed on bloodmeals containing dengue virus as well as controls fed on virus-free control meals at day 1 and day 5 post-feeding. We confirmed infection of midguts from mosquitoes sampled on day 5 post-feeding via RT-PCR. RNAseq analysis revealed dynamic modulation of the expression of several putative immunity and dengue virus-responsive genes, some of whose expression was verified by qRT-PCR. For example, a serine protease gene was up-regulated in the midgut at 1 day post infection, which may potentially enhance mosquito susceptibility to dengue infection, while 14 leucine-rich repeat genes, previously shown to be involved in mosquito antiviral defenses, were down-regulated in the carcass at 5 days post infection. The number of significantly modulated genes decreased over time in midguts and increased in carcasses.

CONCLUSION/SIGNIFICANCE: Dengue virus exposure results in the modulation of genes in a time- and site-specific manner. Previous literature on the interaction between mosquitoes and mosquito-borne pathogens suggests that most of the changes that occurred in Ae. albopictus exposed to DENV would favor virus infection. Many genes identified in this study warrant further characterization to understand their role in viral manipulation of and antiviral response of Ae. albopictus.

摘要

背景

白纹伊蚊,即亚洲虎蚊,目前是登革热、基孔肯雅热和寨卡病毒的重要传播媒介,由于其能够在温带地区定殖,其在节肢动物传播病毒(虫媒病毒)传播中的作用在未来可能会增强。与登革热、基孔肯雅热和寨卡病毒的主要传播媒介埃及伊蚊不同,白纹伊蚊感染虫媒病毒后的基因反应尚未得到充分表征。在此,我们展示了一项关于白纹伊蚊通过摄取人工血餐暴露于2型登革病毒后转录本表达变化的研究。

方法/主要发现:我们在喂食后第1天和第5天分离了摄取含登革病毒血餐的白纹伊蚊的中肠和无中肠尸体,以及摄取无病毒对照餐的对照样本。我们通过逆转录聚合酶链反应(RT-PCR)确认了喂食后第5天采集的蚊子中肠的感染情况。RNA测序分析揭示了几个假定的免疫和登革病毒反应基因表达的动态调节,其中一些基因的表达通过定量逆转录聚合酶链反应(qRT-PCR)得到了验证。例如,一个丝氨酸蛋白酶基因在感染后第1天在中肠中上调,这可能会增强蚊子对登革热感染的易感性,而14个富含亮氨酸的重复基因,先前已证明参与蚊子的抗病毒防御,在感染后第5天在尸体中下调。在中肠中,显著调节基因的数量随时间减少,而在尸体中则增加。

结论/意义:登革病毒暴露导致基因以时间和位点特异性方式调节。先前关于蚊子与蚊媒病原体相互作用的文献表明,暴露于登革病毒的白纹伊蚊发生的大多数变化都有利于病毒感染。本研究中鉴定的许多基因值得进一步表征,以了解它们在白纹伊蚊的病毒操纵和抗病毒反应中的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/55dd/5289563/ff6d6c4d9ff9/pone.0171345.g001.jpg

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