Rabies-viruses-based retrograde tracers can spread across multiple synapses in a retrograde direction in the nervous system of rodents and primates,making them powerful tools for determining the structure and function...Rabies-viruses-based retrograde tracers can spread across multiple synapses in a retrograde direction in the nervous system of rodents and primates,making them powerful tools for determining the structure and function of the complicated neural circuits of the brain.However,they have some limitations,such as posing high risks to human health and the inability to retrograde trans-synaptic label inputs from genetically-de¯ned starter neurons.Here,we established a new retrograde trans-multi-synaptic tracing method through brain-wide rabies virus glycoprotein(RVG)compensation,followed by glycoprotein-deleted rabies virus(RV-△G)infection in specific brain regions.Furthermore,in combination with the avian tumor virus receptor A(TVA)controlled by a cell-type-specific promoter,we found that EnvA-pseudotyped RV-△G can mediate e±cient retrograde trans-multi-synaptic transduction from cell-type-specific starter neurons.This study provides new alternative methods for neuroscience researchers to analyze the input neural networks of rodents and nonhuman primates.展开更多
Efficient viral vectors for mapping and manipulating long-projection neuronal circuits are crucial in structural and functional studies of the brain. The SAD strain rabies virus with the glycoprotein gene deleted pseu...Efficient viral vectors for mapping and manipulating long-projection neuronal circuits are crucial in structural and functional studies of the brain. The SAD strain rabies virus with the glycoprotein gene deleted pseudotyped with the N2 C glycoprotein(SAD-RV(DG)-N2 C(G)) shows strong neuro-tropism in cell culture, but its in vivo efficiency for retrograde gene transduction and neuro-tropism have not been systematically characterized.We compared these features in different mouse brain regions for SAD-RV-N2 C(G) and two other widely-used retrograde tracers, SAD-RV(DG)-B19(G) and r AAV2-retro. We found that SAD-RV(DG)-N2 C(G) enhanced the infection efficiency of long-projecting neurons by^10 times but with very similar neuro-tropism, compared with SAD-RV(DG)-B19(G). On the other hand, SAD-RV(DG)-N2 C(G) had an infection efficiency comparable with r AAV2-retro, but a more restricted diffusion range, and broader tropism to different types and regions of longprojecting neuronal populations. These results demonstrate that SAD-RV(DG)-N2 C(G) can serve as an effective retrograde vector for studying neuronal circuits.展开更多
基金study was supported by the STI2030-Major Projects(Grant No.2021ZD0201003)the National Natural Science Foundation of China(Grant Nos.31830035,31771156,21921004,and 32100899)+3 种基金the Strategic Priority Research Program of the Chinese Academy of Sciences(Grant No.XDB32030200)the Shenzhen Key Laboratory of Viral Vectors for Biomedicine(Grant No.ZDSYS20200811142401005)the Key Laboratory of Quality Control Technology for Virus-Based Ther-apeutics,Guangdong Provincial Medical Products Administration(Grant No.2022ZDZ13)Open Project Program of Wuhan National Laboratory for Optoelectronics(Grant No.2019WNLOKF022).
文摘Rabies-viruses-based retrograde tracers can spread across multiple synapses in a retrograde direction in the nervous system of rodents and primates,making them powerful tools for determining the structure and function of the complicated neural circuits of the brain.However,they have some limitations,such as posing high risks to human health and the inability to retrograde trans-synaptic label inputs from genetically-de¯ned starter neurons.Here,we established a new retrograde trans-multi-synaptic tracing method through brain-wide rabies virus glycoprotein(RVG)compensation,followed by glycoprotein-deleted rabies virus(RV-△G)infection in specific brain regions.Furthermore,in combination with the avian tumor virus receptor A(TVA)controlled by a cell-type-specific promoter,we found that EnvA-pseudotyped RV-△G can mediate e±cient retrograde trans-multi-synaptic transduction from cell-type-specific starter neurons.This study provides new alternative methods for neuroscience researchers to analyze the input neural networks of rodents and nonhuman primates.
基金This work was supported by the National Basic Research Program of China(2017YFA0103303)Strategic Priority Research Program of the Chinese Academy of Sciences(XDB32010100,XDB02050400,XDB02050005,XDA16020601)+2 种基金National Basic Research Program of China(2017YFA0102601,2019YFA0110100)National Natural Science Foundation of China(NSFC)(91732301,31671072,31771140,81891001,91432111,81527901,31400977,31625013)Grants of Beijing Brain Initiative of Beijing Municipal Science&Technology Commission(Z181100001518004).
基金the National Basic Research Program(973 Program)of China(2015CB755601)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDB32030200)+1 种基金the National Natural Science Foundation of China(31771156,81661148053,91632303,31800885,31500868,31671120 and 91732304)the China Postdoctoral Science Foundation(2019M653118 and 2018M632946).
文摘Efficient viral vectors for mapping and manipulating long-projection neuronal circuits are crucial in structural and functional studies of the brain. The SAD strain rabies virus with the glycoprotein gene deleted pseudotyped with the N2 C glycoprotein(SAD-RV(DG)-N2 C(G)) shows strong neuro-tropism in cell culture, but its in vivo efficiency for retrograde gene transduction and neuro-tropism have not been systematically characterized.We compared these features in different mouse brain regions for SAD-RV-N2 C(G) and two other widely-used retrograde tracers, SAD-RV(DG)-B19(G) and r AAV2-retro. We found that SAD-RV(DG)-N2 C(G) enhanced the infection efficiency of long-projecting neurons by^10 times but with very similar neuro-tropism, compared with SAD-RV(DG)-B19(G). On the other hand, SAD-RV(DG)-N2 C(G) had an infection efficiency comparable with r AAV2-retro, but a more restricted diffusion range, and broader tropism to different types and regions of longprojecting neuronal populations. These results demonstrate that SAD-RV(DG)-N2 C(G) can serve as an effective retrograde vector for studying neuronal circuits.
