Drosophila melanogaster as a model organism for bluetongue virus replication and tropism.
Drosophila melanogaster as a model organism for bluetongue virus replication and tropism.
复制标题
果蝇作为蓝舌病毒复制和向性的模式生物。
DOI：
10.1128/jvi.00131-12
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发表时间：
2012
影响因子：
5.4
通讯作者：
Shaw AE
中科院分区：
文献类型：
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作者：
Shaw AE
蓝舌病毒(BTV)是蓝舌病(Bt)的病原体，蓝舌病是反刍动物的一种出血性疾病，可导致高水平的发病率和死亡率。BTV是一种虫媒病毒，通过库蚊叮咬蚊子在其反刍动物宿主之间传播。最近，欧洲经历了一些有记录以来最大规模的Bt疫情，包括一些没有已知病史的地区，导致了前所未有的经济和动物福利问题。目前缺乏库蚊基因组资源和遗传工具，限制了对病毒与昆虫相互作用机制的详细研究。相比之下，果蝇的基因组已经成功测序，由于强大的基因技术的存在，它被广泛用作分子途径的模型。在这项研究中，黑腹夜蛾被作为BTV复制和趋向性的模型进行研究。利用反向遗传学技术，构建了表达荧光mCherry蛋白与病毒非结构蛋白NS3融合的改良BTV-1(BTV-1/NS3mCherry)。我们证明了BTV-1/NS3mCherry不仅具有复制能力，因为它保留了野生型病毒的许多特征，而且在通过抗生素治疗去除细菌内共生菌Wolbachia piientis后，还可以在黑腹葡萄球菌中有效复制。此外，共聚焦显微镜显示，BTV-1/NS3mCherry在黑腹夜蛾中的组织趋向性与先前在库蚊中描述的BTV相似。总体而言，这项研究中提供的数据表明，使用黑腹夜蛾作为遗传模型来研究BTV与昆虫的相互作用是可行的，否则无法在媒介物种中解决这一问题。
Bluetongue virus (BTV) is the etiological agent of bluetongue (BT), a hemorrhagic disease of ruminants that can cause high levels of morbidity and mortality. BTV is an arbovirus transmitted between its ruminant hosts by Culicoides biting midges (Diptera: Ceratopogonidae). Recently, Europe has experienced some of the largest BT outbreaks ever recorded, including areas with no known history of the disease, leading to unprecedented economic and animal welfare issues. The current lack of genomic resources and genetic tools for Culicoides restricts any detailed study of the mechanisms involved in the virus-insect interactions. In contrast, the genome of the fruit fly (Drosophila melanogaster) has been successfully sequenced, and it is used extensively as a model of molecular pathways due to the existence of powerful genetic technology. In this study, D. melanogaster is investigated as a model for the replication and tropism of BTV. Using reverse genetics, a modified BTV-1 that expresses the fluorescent mCherry protein fused to the viral nonstructural protein NS3 (BTV-1/NS3mCherry) was generated. We demonstrate that BTV-1/NS3mCherry is not only replication competent as it retains many characteristics of the wild-type virus but also replicates efficiently in D. melanogaster after removal of the bacterial endosymbiont Wolbachia pipientis by antibiotic treatment. Furthermore, confocal microscopy shows that the tissue tropism of BTV-1/NS3mCherry in D. melanogaster resembles that described previously for BTV in Culicoides. Overall, the data presented in this study demonstrate the feasibility of using D. melanogaster as a genetic model to investigate BTV-insect interactions that cannot be otherwise addressed in vector species.