Axonal transport and sorting of herpes simplex virus components in a mature mouse visual system
Axonal transport and sorting of herpes simplex virus components in a mature mouse visual system
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DOI：
10.1128/jvi.77.11.6117-6126.2003
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发表时间：
2003-06-01
影响因子：
5.4
通讯作者：
Sidhu, SS
中科院分区：
文献类型：
--
作者：
LaVail, JH;Tauscher, AN;Sidhu, SS
研究了单纯疱疹病毒（HSV）两种主要蛋白在成熟小鼠视网膜神经节细胞轴突体内的传递和转运过程。在玻璃体内注射HSV 24小时后，将伐昔洛韦引入小鼠的饮用水中，以抑制随后的病毒复制。未经治疗，病毒在轴突周围胶质细胞中的传播和复制混淆了轴突运输的研究。感染后2至5天，处死动物，切除视神经通路的连续片段。免疫荧光显微镜显示感染的星形胶质细胞数量在视神经近端减少，在视神经束中消失。用包膜和衣壳成分抗体、糖蛋白D （gD）和VP5抗体对视网膜进行Western blot检测，结果显示这两种成分在视网膜匀浆中表达2天。逆转录pcr结果显示，感染后5天视神经束内不存在gD mRNA。因此，我们认为gD是由视网膜神经节细胞体中的病毒mRNA转录而来。gD在近节细胞轴突积累2天，3天后到达最远节段。VP5在gD出现后约48 h，即第4天首次出现在近端轴突。因此，gD较早进入轴突，且不依赖于VP5。这些发现证实了病毒在神经元中转运的亚组装模型，并表明使用valacyclovir进行有效抗病毒治疗有4- 5天的窗口期。
The time course for delivery and transport of two major proteins of herpes simplex virus (HSV) has been determined for mature mouse retinal ganglion cell axons in vivo. Twenty-four hours after intravitreal injection of HSV, valacyclovir was introduced into the drinking water of the mice to inhibit subsequent viral replication. Without treatment, viral spread and replication in periaxonal glial cells confound study of axonal transport. At 2 to 5 days after infection, the animals were sacrificed and contiguous segments of the optic pathway were removed. Immunofluorescence microscopy indicated that the number of infected astrocytes was reduced in the proximal optic nerve and eliminated in the optic tract. Western blots of the retina with antibodies for envelope and capsid components, glycoprotein D (gD) and VP5, respectively, revealed that both components were expressed in retinal homogenates by 2 days. Results of reverse transcription-PCR indicated that there was no gD mRNA present in the treated optic tract 5 days after infection. Therefore, we conclude that gD is transcribed from viral mRNA in the retinal ganglion cell bodies. The gD accumulated in the proximal ganglion cell axon by 2 days and reached the most distal segment after 3 days. The VP5 first appeared in the proximal axons at 4 days, about 48 h after the appearance of gD. Thus, gD entered the axon earlier and independent of VP5. These finding confirm the subassembly model of viral transport in neurons and suggest that there is a 4- to 5-day window for initiation of effective antiviral treatment with valacyclovir.