Resolving the Assembly State of Herpes Simplex Virus during Axon Transport by Live-Cell Imaging
Resolving the Assembly State of Herpes Simplex Virus during Axon Transport by Live-Cell Imaging
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DOI：
10.1128/jvi.01296-10
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发表时间：
2010-12-01
影响因子：
5.4
通讯作者：
Smith, Gregory A.
中科院分区：
文献类型：
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作者：
Antinone, Sarah E.;Zaichick, Sofia V.;Smith, Gregory A.
嗜神经疱疹病毒依赖于远距离轴突运输在外周神经节初始潜伏期的建立（逆行运输）和病毒在再激活后轴突向暴露体表的传播（顺行运输）。使用电子显微镜获得的1型单纯疱疹病毒（HSV-1）感染神经元的图像引发了关于为什么在轴突中看到不同类型的病毒结构以及这些颗粒中哪些与轴突运输过程有关的争论。在这项研究中，我们应用延时荧光显微镜对原代神经元和神经元细胞系中主动转移到远端轴突的HSV-1病毒粒子组分进行了成像。这些发现的关键是，在任何给定时间，只有一小部分病毒颗粒在感染的出口阶段进行顺行运输。通过对主动转运衣壳亚群组成的选择性分析，确定了完全组装的HSV-1病毒粒子的运输与神经元分泌途径之间的联系。最后，我们评估了先前在固定细胞中HSV-1轴突运输的研究中看似相反的发现，并证明了使用抗体检测方法评估单个HSV-1颗粒组成的局限性。
Neurotropic herpesviruses depend on long-distance axon transport for the initial establishment of latency in peripheral ganglia (retrograde transport) and for viral spread in axons to exposed body surfaces following reactivation (anterograde transport). Images of neurons infected with herpes simplex virus type 1 (HSV-1), acquired using electron microscopy, have led to a debate regarding why different types of viral structures are seen in axons and which of these particles are relevant to the axon transport process. In this study, we applied time-lapse fluorescence microscopy to image HSV-1 virion components actively translocating to distal axons in primary neurons and neuronal cell lines. Key to these findings, only a small fraction of viral particles were engaged in anterograde transport during the egress phase of infection at any given time. By selective analysis of the composition of the subpopulation of actively transporting capsids, a link between transport of fully assembled HSV-1 virions and the neuronal secretory pathway was identified. Last, we have evaluated the seemingly opposing findings made in previous studies of HSV-1 axon transport in fixed cells and demonstrate a limitation to assessing the composition of individual HSV-1 particles using antibody detection methods.