Trans-synaptic Neural Circuit-Tracing with Neurotropic Viruses

Li, Jiamin; Liu, Taian; Dong, Yun; Kondoh, Kunio; Lu, Zhonghua

doi:10.1007/s12264-019-00374-9

Cited by 50 publications

(31 citation statements)

References 111 publications

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“…Various neurotropic viruses, such as HSV1 [22], CAV2 [6], RABV [13], HiRet or NeuRet lentivirus [25], which have been widely used in the study of brain circuits, have the characteristics of retrograde access to speci c or multiple types of neurons [63,64]. However, high toxicity made them not conducive to longterm functional studies in model animals, which further limited their application in disease treatment [4].…”

Section: Discussionmentioning

confidence: 99%

AAV9-Retro Mediates Efficient Transduction with Axon Terminal Absorption and Blood-brain Barrier Transportation

Lin

Zhong

et al. 2020

Preprint

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Recombinant adeno-associated viruses (rAAVs), especially which permit efficient gene transfer to neurons from axonal terminals or across the blood-brain barrier, are useful vehicles for the structural and functional studies of neural circuit, and for the treatment of many gene-deficient brain diseases that needs to compensate for the correct genes to every cell in the whole brain. However, AAVs with these two advantages have not been reported. Here, we describe a new capsid engineering method, which draws on advantage combination of different capsids, and aims to yield a capsid that can provide more alternative routes of administration, which are more suitable for wide-scale transduction of the CNS. A new AAV variant, AAV9-Retro, was developed by inserting the 10-mer peptide fragment from AAV2-Retro into the capsid of AAV9, and the biodistribution properties were evaluated in mice. By intracranial and intravenous injection in the mice, we found that AAV9-Retro can retrogradely infect projection neurons with efficiency comparable to AAV2-Retro, and retains the characteristic of AAV9 that can transport across the nervous system. Our strategy provides a new tool for the manipulation of neural circuits, and for the future preclinical and clinical treatment of some neurological and neurodegenerative disorders.

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Section: Discussionmentioning

confidence: 99%

AAV9-Retro Mediates Efficient Transduction with Axon Terminal Absorption and Blood-brain Barrier Transportation

Lin

Zhong

et al. 2020

Preprint

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“…Neurotropic viruses have the innate ability to invade neurons and produce infectious progeny that spread along synaptically connected neuronal networks [1][2][3]. Mapping neural circuits requires both anterograde and retrograde tracers [4]. To date, rabies virus and pseudorabies virus (PRV) strains have been modified for use as retrograde tracers [5][6][7][8].…”

Section: Introductionmentioning

confidence: 99%

High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy

Ying

Han

et al. 2020

Mol Brain

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Neurotropic viral transsynaptic tracing is an increasingly powerful technique for dissecting the structure and function of neural circuits. Herpes simplex virus type 1 strain H129 has been widely used as an anterograde tracer. However, HSV tracers still have several shortcomings, including high toxicity, low sensitivity and non-specific retrograde labeling. Here, we aimed to construct high-brightness HSV anterograde tracers by increasing the expression of exogenous genes carried by H129 viruses. Using a Trojan horse-like strategy, a HSV/AAV (adenoassociated virus) chimaera termed H8 was generated to enhance the expression of a fluorescent marker. In vitro and in vivo assays showed that the exogenous gene was efficiently replicated and amplified by the synergism of the HSV vector and introduced AAV replication system. H8 reporting fluorescence was brighter than that of currently available H129 tracers, and H8 could be used for fast and effective anterograde tracing without additional immunostaining. These results indicated that foreign gene expression in HSV tracers could be enhanced by integrating HSV with AAV replication system. This approach may be useful as a general enhanced expression strategy for HSV-based tracing tools or gene delivery vectors.

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“…Envelope glycoprotein of VSV can be replaced by glycoproteins of other viruses to achieve different labeling characteristics and direction controllable trans-synaptic transfer [21,22]. But these VSVs have high cytotoxicity that can lead to rapid death of mice, which limits long-term structural observation and gene manipulation or function research, restricting their applications in neuroscience [23]. Several mutants have been obtained by mutating the N gene related to the replication of VSV [24][25][26][27].…”

Section: Introductionmentioning

confidence: 99%

A mutant vesicular stomatitis virus with reduced cytotoxicity and enhanced anterograde trans-synaptic efficiency

Lin,

Zhong,

Ying

et al. 2020

Mol Brain

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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-N R7A , and examined its cytotoxicity and efficiency of trans-synaptic spreading. We found that by comparison with the wild-type tracer of VSV, the N R7A mutation 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-N R7A , 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.

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Trans-synaptic Neural Circuit-Tracing with Neurotropic Viruses

Cited by 50 publications

References 111 publications

AAV9-Retro Mediates Efficient Transduction with Axon Terminal Absorption and Blood-brain Barrier Transportation

AAV9-Retro Mediates Efficient Transduction with Axon Terminal Absorption and Blood-brain Barrier Transportation

High-brightness anterograde transneuronal HSV1 H129 tracer modified using a Trojan horse-like strategy

A mutant vesicular stomatitis virus with reduced cytotoxicity and enhanced anterograde trans-synaptic efficiency

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