中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2022, Vol. 17 ›› Issue (12): 2761-2764.doi: 10.4103/1673-5374.339495

• 原著:神经再生技术 • 上一篇    下一篇

基于辛德毕斯病毒的新型顺向跨突触示踪工具

  

  • 出版日期:2022-12-15 发布日期:2022-05-05
  • 基金资助:
    国家自然科学基金项目（31830035，91732304，91632303，81661148053，31771156）；广东省重点领域研究发展计划项目（2018B030331001）；SIAT中国优秀青年科学家创新计划项目（E1G023）；广东省基础与应用基础研究基金项目（2021A1515011235）；深圳市生物医学病毒载体重点实验室项目（ZDSYS20200811142401005）；国家基础研究计划（973计划）项目（2015CB755600）；国家战略性先导研究计划项目（B类）（XDB32030200）

A new anterograde trans-synaptic tracer based on Sindbis virus

Xiang-Wei Shi1, 2, 3, 4, Fan Jia2, 3, 4, *, Pei Lyu1, Fu-Qiang Xu1, 2, 3, 4, 5, *   

  1. 1State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Key Laboratory of Magnetic Resonance in Biological Systems, Wuhan Center for Magnetic Resonance, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan, Hubei Province, China; 2Guangdong Provincial Key Laboratory of Brain Connectome and Behavior, CAS Key Laboratory of Brain Connectome and Manipulation, the Brain Cognition and Brain Disease Institute (BCBDI), Translational Research Center for the Nervous System (TRCNS), Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences; Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research Institutions, Shenzhen, Guangdong Province, China; 3NMPA Key Laboratory for Research and Evaluation of Viral Vector Technology in Cell and Gene Therapy Medicinal Products, Key Laboratory of Quality Control Technology for Virus-Based Therapeutics, Guangdong Provincial Medical Products Administration, Shenzhen Key Laboratory of Viral Vectors for Biomedicine, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen, Guangdong Province, China; 4University of Chinese Academy of Sciences, Beijing, China; 5Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology, Wuhan, Hubei Province, China
  • Online:2022-12-15 Published:2022-05-05
  • Contact: Fu-Qiang Xu, PhD, fq.xu@siat.ac.cn; Fan Jia, PhD, fan.jia@siat.ac.cn.
  • Supported by:
    This work was supported by the National Natural Science Foundation of China, Nos. 31830035, 91732304, 91632303, 81661148053, and 31771156 (all to FQX); the Key-Area Research and Development Program of Guangdong Province of China, No. 2018B030331001 (to FQX); the SIAT Innovation Program for Excellent Young Researchers of China, No. E1G023 (to FJ); the Guangdong Basic and Applied Basic Research Foundation of China, No. 2021A1515011235 (to FQX); Shenzhen Key Laboratory of Viral Vectors for Biomedicine of China, No. ZDSYS20200811142401005 (to FQX); the National Basic Research Program (973 Program) of China, No. 2015CB755600 (to FQX); and the Strategic Priority Research Program (B) of China, No. XDB32030200 (to FQX).

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摘要：

绘制神经回路对于了解神经系统的结构和功能至关重要，工程化病毒是一种示踪神经回路的有价值方法。然而现有的示踪工具不能满足实验需求，因而急需开发新型的示踪工具。实验构建了一种可表达增强型绿色荧光蛋白的工程化辛德毕斯病毒，并发现其与野生型辛德毕斯病毒在感染BHK-21细胞和体外神经元方面无明显差异。然后将表达增强型绿色荧光蛋白的工程化辛德毕斯病毒注射到小鼠大脑视觉通路中，发现病毒可感染注射部位的神经元，并在神经回路中顺行扩散。因此表达增强型绿色荧光蛋白的工程化辛德毕斯病毒可作为一种新型顺向示踪工具，用于标记及示踪脑神经环路。

https://orcid.org/0000-0002-7779-5556 (Fan Jia); https://orcid.org/0000-0003-0711-2147 (Xiang-Wei Shi)

关键词: 辛德毕斯病毒, 顺行, 示踪剂, 突触, 神经回路, 小鼠大脑, 神经元, 视网膜, 外侧膝状体, 上丘

Abstract: Mapping neural circuits is critical for understanding the structure and function of the nervous system. Engineered viruses are a valuable tool for tracing neural circuits. However, current tracers do not fully meet the needs for this approach because of various drawbacks, such as toxicity and characteristics that are difficult to modify. Therefore, there is an urgent need to develop a new tracer with low toxicity and that allows for long-term studies. In this study, we constructed an engineered Sindbis virus (SINV) expressing enhanced green fluorescent protein (EGFP) reporter gene (SINV-EGFP) and found that it had no significant difference in biological characterization compared with the wild-type Sindbis virus in BHK-21 cells and neurons in vitro. We injected the virus into the visual circuit of mouse brain and found that the virus infected neurons in the local injected site and anterogradely spread in the neural circuits. Although the efficiency of transmission was limited, the findings demonstrate that SINV can be used as a new anterograde tracer to map neural circuits in mouse brain and that it spreads exclusively in the anterograde direction. Further, use of SINV in mouse brain research will provide longer time windows for circuit tracing than is possible with herpes simplex virus and vesicular stomatitis virus tracers.

Key words: anterograde, lateral geniculate nucleus, mouse brains, neural circuit, neurons, retina, Sindbis virus, superior colliculus, synapse, tracer