Retrograde axonal transport of herpes simplex virus: Evidence for a single mechanism and a role for tegument
Retrograde axonal transport of herpes simplex virus: Evidence for a single mechanism and a role for tegument
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DOI：
10.1073/pnas.97.14.8146
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发表时间：
2000-07-05
影响因子：
11.1
通讯作者：
LaVail, JH
中科院分区：
文献类型：
--
作者：
Bearer, EL;Breakefield, XO;LaVail, JH
I型单纯疱疹病毒（HSV）通常进入周围神经末梢，然后沿着神经返回到达神经元细胞体，在那里复制或进入潜伏期。为了监测HSV的轴质运输，我们使用了鱿鱼的巨大轴突Loligo pealei，这是一个众所周知的轴质运输研究系统。为了将HSV传递到轴质中，用洗涤剂剥离包膜的病毒颗粒被注射到巨大的轴突中，从而绕过感染过程。用绿色荧光蛋白标记病毒被膜蛋白VP16，可以用共聚焦显微镜对在轴突内移动的病毒颗粒进行成像。病毒颗粒在逆行方向上移动2.2 +/- 0.26 μ m/秒，这一速度与内源性细胞器和病毒在培养的哺乳动物神经元中的运输速度相当。电子显微镜证实96%的运动（剥离）病毒颗粒失去了包膜，但保留了被膜，Western blot分析显示这些颗粒保留了衣壳的蛋白质，但没有包膜。我们得出结论：(i) HSV招募鱿鱼逆行运输机械；（ii）病毒外皮和衣壳（而非包膜）足以进行这种招募；（iii）鱿鱼巨大的轴突提供了一个独特的系统来解剖运输所需的病毒成分，并确定它们招募的细胞运输机制。
Herpes simplex virus type I (HSV) typically enters peripheral nerve terminals and then travels back along the nerve to reach the neuronal cell body, where it replicates or enters latency. To monitor axoplasmic transport of HSV, we used the giant axon of the squid, Loligo pealei, a well known system for the study of axoplasmic transport. To deliver HSV into the axoplasm, viral particles stripped of their envelopes by detergent were injected into the giant axon, thereby bypassing the infective process. Labeling the viral tegument protein, VP16, with green fluorescent protein allowed viral particles moving inside the axon to be imaged by confocal microscopy. Viral particles moved 2.2 +/- 0.26 mu m/sec in the retrograde direction, a rate comparable to that of the transport of endogenous organelles and of virus in mammalian neurons in culture. Electron microscopy confirmed that 96% of motile (stripped) viral particles had lost their envelope but retained tegument, and Western blot analysis revealed that these particles had retained protein from capsid but not envelope. We conclude that (i) HSV recruits the squid retrograde transport machinery; (ii) viral tegument and capsid but not envelope are sufficient for this recruitment; and (iii) the giant axon of the squid provides a unique system to dissect the viral components required for transport and to identify the cellular transport mechanisms they recruit.