A rabies virus-based toolkit for efficient retrograde labeling and monosynaptic tracing

作     者：Kun-Zhang Lin Lei Li Wen-Yu Ma Xin Yang Zeng-Peng Han Neng-Song Luo Jie Wang Fu-Qiang Xu Kun-Zhang Lin;Lei Li;Wen-Yu Ma;Xin Yang;Zeng-Peng Han;Neng-Song Luo;Jie Wang;Fu-Qiang Xu

作者机构：The Brain Cognition and Brain Disease Institute(BCBDI)Shenzhen Key Laboratory of Viral Vectors for BiomedicineShenzhen Institute of Advanced TechnologyChinese Academy of SciencesShenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research InstitutionsNMPA Key Laboratory for Research and Evaluation of Viral Vector Technology in Cell and Gene Therapy Medicinal ProductsShenzhenKey Laboratory of Quality Control Technology for Virus-Based TherapeuticsGuangdong Provincial Medical Products AdministrationShenzhenGuangdong ProvinceChina State Key Laboratory of Magnetic Resonance and Atomic and Molecular PhysicsNational Center for Magnetic Resonance in WuhanWuhan Institute of Physics and MathematicsInnovation Academy for Precision Measurement Science and TechnologyChinese Academy of Sciences-Wuhan National Laboratory for OptoelectronicsWuhanHubei ProvinceChina University of Chinese Academy of SciencesBeijingChina Wuhan National Laboratory for OptoelectronicsHuazhong University of Science and TechnologyWuhanHubei ProvinceChina Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research InstitutionsShenzhenGuangdong ProvinceChina Center for Excellence in Brain Science and Intelligence TechnologyChinese Academy of SciencesShanghaiChina 

出 版 物：《Neural Regeneration Research》 (中国神经再生研究（英文版）)

年 卷 期：2023年第18卷第8期

页      面：1827-1833页

核心收录：

学科分类：0710[理学-生物学] 1001[医学-基础医学(可授医学、理学学位)] 100103[医学-病原生物学] 10[医学] 

基　　金：supported by the National Natural Science Foundation of China,Nos.32100899(to KZL),31830035(to FQX),31771156(to FQX),21921004(to FQX) the National Science and Technology Innovation 2030,No.2021ZD0201003(to FQX) the Key-Area Research and Development Program of Guangdong Province,No.2018B030331001(to FQX) the Strategic Priority Research Program of the Chinese Academy of Sciences,No.XDB32030200(to FQX) the Shenzhen Key Laboratory of Viral Vectors for Biomedicine,No.ZDSYS20200811142401005(to FQX)。 

主　　题：functional studies neural activity neural circuits projection neurons rAAV9-Retro rabies virus recombination retrograde labeling synaptic inputs trans-monosynaptic tracing 

摘      要：Analyzing the structure and function of the brain s neural network is critical for identifying the working principles of the brain and the mechanisms of brain diseases.Recombinant rabies viral vectors allow for the retrograde labeling of projection neurons and cell type-specific trans-monosynaptic tracing,making these vectors powerful candidates for the dissection of synaptic inputs.Although several attenuated rabies viral vectors have been developed,their application in studies of functional networks is hindered by the long preparation cycle and low yield of these vectors.To overcome these limitations,we developed an improved production system for the rapid rescue and preparation of a high-titer CVS-N2c-ΔG virus.Our results showed that the new CVS-N2c-ΔG-based toolkit performed remarkably:(1)N2cG-coated CVS-N2c-ΔG allowed for efficient retrograde access to projection neurons that were unaddressed by rAAV9-Retro,and the efficiency was six times higher than that of rAAV9-Retro;(2)the trans-monosynaptic efficiency of oG-mediated CVS-N2c-ΔG was 2–3 times higher than that of oG-mediated SAD-B19-ΔG;(3)CVS-N2c-ΔG could delivery modified genes for neural activity monitoring,and the time window during which this was maintained was 3 weeks;and(4)CVS-N2c-ΔG could express sufficient recombinases for efficient transgene recombination.These findings demonstrate that new CVS-N2c-ΔG-based toolkit may serve as a versatile tool for structural and functional studies of neural circuits.