A mutant vesicular stomatitis virus with reduced cytotoxicity and enhanced anterograde trans-synaptic efficiency
A mutant vesicular stomatitis virus with reduced cytotoxicity and enhanced anterograde trans-synaptic efficiency
复制标题
一种突变型水泡性口炎病毒，具有降低的细胞毒性和增强的顺行跨突触效率
DOI：
10.1186/s13041-020-00588-3
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发表时间：
2020-03-20
期刊：
影响因子：
3.6
通讯作者：
Xu, Fuqiang
中科院分区：
文献类型：
--
作者：
Lin, Kunzhang;Zhong, Xin;Xu, Fuqiang
了解脑网络的连接结构是揭示脑功能原理和阐明脑疾病发生机制的基础。嗜神经病毒跨突触示踪技术已成为研究神经回路的最有效的技术之一。尽管利用重组狂犬病病毒和伪狂犬病病毒分析输入神经网络的逆行跨突触追踪方法在神经科学中得到了广泛的应用，但用于分析输出神经网络的病毒工具仍然缺乏。重组水泡性口炎病毒（VSV）已被用于映射突触输出。然而，一些缺点，包括高神经毒性和实验动物的快速致死性，阻碍了其在神经网络结构和功能的长期研究中的应用。为了克服这些局限性，我们产生了一个重组VSV复制相关的N基因突变，VSV-NR 7A，并检查其细胞毒性和跨突触传播的效率。我们发现，与野生型示踪剂VSV相比，NR 7A突变赋予病毒较低的增殖率和体外细胞毒性，以及显着减少体内神经炎症反应和更长的动物存活时间，当它被注射到小鼠脑细胞核中。此外，VSV减毒后注入腹侧被盖区可延迟其扩散。重要的是，随着毒性的降低和动物存活时间的延长，突变VSV跨突触标记的脑区域的数量多于野生型VSV。这些结果表明，VSV-NR 7A可能是一种有前途的顺行示踪剂，使研究人员能够探索给定脑区的更多下游连接，并在更长的时间窗口内观察下游回路的解剖结构和功能。我们的工作为神经回路的结构和功能研究提供了一个更好的工具。
Understanding the connecting structure of brain network is the basis to reveal the principle of the brain function and elucidate the mechanism of brain diseases. Trans-synaptic tracing with neurotropic viruses has become one of the most effective technologies to dissect the neural circuits. Although the retrograde trans-synaptic tracing for analyzing the input neural networks with recombinant rabies and pseudorabies virus has been broadly applied in neuroscience, viral tools for analyzing the output neural networks are still lacking. The recombinant vesicular stomatitis virus (VSV) has been used for the mapping of synaptic outputs. However, several drawbacks, including high neurotoxicity and rapid lethality in experimental animals, hinder its application in long-term studies of the structure and function of neural networks. To overcome these limitations, we generated a recombinant VSV with replication-related N gene mutation, VSV-NR7A, and examined its cytotoxicity and efficiency of trans-synaptic spreading. We found that by comparison with the wild-type tracer of VSV, the NR7Amutation endowed the virus lower rate of propagation and cytotoxicity in vitro, as well as significantly reduced neural inflammatory responses in vivo and much longer animal survival when it was injected into the nucleus of the mice brain. Besides, the spreading of the attenuated VSV was delayed when injected into the VTA. Importantly, with the reduced toxicity and extended animal survival, the number of brain regions that was trans-synaptically labeled by the mutant VSV was more than that of the wild-type VSV. These results indicated that the VSV-NR7A, could be a promising anterograde tracer that enables researchers to explore more downstream connections of a given brain region, and observe the anatomical structure and the function of the downstream circuits over a longer time window. Our work could provide an improved tool for structural and functional studies of neurocircuit.