An HSV-1-H129 amplicon tracer system for rapid and efficient monosynaptic anterograde neural circuit tracing

Feng Xiong; Hong Yang; Yi-Ge Song; Hai-Bin Qin; Qing-Yang Zhang; Xian Huang; Wei Jing; Manfei Deng; Yang Liu; Zhixiang Liu; Yin Shen; Yunyun Han; Youming Lu; Xiangmin Xu; Todd C Holmes; Minmin Luo; Fei Zhao; Min-Hua Luo; Wen-Bo Zeng

doi:10.1038/s41467-022-35355-6

An HSV-1-H129 amplicon tracer system for rapid and efficient monosynaptic anterograde neural circuit tracing

Nat Commun. 2022 Dec 10;13(1):7645. doi: 10.1038/s41467-022-35355-6.

Authors

Feng Xiong^#^{1

2

3}, Hong Yang^#^{1

2}, Yi-Ge Song⁴, Hai-Bin Qin^{1

2}, Qing-Yang Zhang^{1

2}, Xian Huang⁴, Wei Jing⁴, Manfei Deng⁴, Yang Liu⁵, Zhixiang Liu⁵, Yin Shen⁶, Yunyun Han⁶, Youming Lu⁴, Xiangmin Xu^{7

8}, Todd C Holmes^{8

9}, Minmin Luo^{5

10}, Fei Zhao^{11

12}, Min-Hua Luo^{13

14

15

16}, Wen-Bo Zeng¹⁷

Affiliations

¹ State Key Laboratory of Virology, CAS Center for Excellence in Brain Science and Intelligence Technology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China.
² University of Chinese Academy of Sciences, Beijing, China.
³ Key Laboratory of Magnetic Resonance in Biological Systems, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy of Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan, China.
⁴ Department of Physiology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
⁵ National Institute of Biological Sciences, Beijing, China.
⁶ Eye Center, Renmin Hospital, Wuhan University, Wuhan, China.
⁷ Department of Anatomy and Neurobiology, School of Medicine, University of California, Irvine, CA, USA.
⁸ Center for Neural Circuit Mapping, School of Medicine, University of California, Irvine, CA, USA.
⁹ Department of Physiology and Biophysics, School of Medicine, University of California, Irvine, CA, USA.
¹⁰ Chinese Institute for Brain Research, Beijing, China.
¹¹ Chinese Institute for Brain Research, Beijing, China. zhaofei@cibr.ac.cn.
¹² School of Basic Medical Sciences, Capital Medical University, Beijing, China. zhaofei@cibr.ac.cn.
¹³ State Key Laboratory of Virology, CAS Center for Excellence in Brain Science and Intelligence Technology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China. luomh@wh.iov.cn.
¹⁴ University of Chinese Academy of Sciences, Beijing, China. luomh@wh.iov.cn.
¹⁵ Key Laboratory of Magnetic Resonance in Biological Systems, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, National Center for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy of Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan, China. luomh@wh.iov.cn.
¹⁶ Center for Neural Circuit Mapping, School of Medicine, University of California, Irvine, CA, USA. luomh@wh.iov.cn.
¹⁷ State Key Laboratory of Virology, CAS Center for Excellence in Brain Science and Intelligence Technology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China. zengwb@wh.iov.cn.

^# Contributed equally.

Abstract

Monosynaptic viral tracers are essential tools for dissecting neuronal connectomes and for targeted delivery of molecular sensors and effectors. Viral toxicity and complex multi-injection protocols are major limiting application barriers. To overcome these barriers, we developed an anterograde monosynaptic H129_Amp tracer system based on HSV-1 strain H129. The H129_Amp tracer system consists of two components: an H129-dTK-T2-pac^Flox helper which assists H129_Amp tracer's propagation and transneuronal monosynaptic transmission. The shared viral features of tracer/helper allow for simultaneous single-injection and subsequent high expression efficiency from multiple-copy of expression cassettes in H129_Amp tracer. These improvements of H129_Amp tracer system shorten experiment duration from 28-day to 5-day for fast-bright monosynaptic tracing. The lack of toxic viral genes in the H129_Amp tracer minimizes toxicity in postsynaptic neurons, thus offering the potential for functional anterograde mapping and long-term tracer delivery of genetic payloads. The H129_Amp tracer system is a powerful tracing tool for revealing neuronal connectomes.

MeSH terms

Connectome*
Herpesvirus 1, Human / genetics
Nerve Net*
Neurons