Optimization of the Fluorescent Protein Expression Level Based on Pseudorabies Virus Bartha Strain for Neural Circuit Tracing

Fan, Jia; Lv, Pei; Miao, Huan; Shi, Xiangwei; Mei, Hongjun; Li, Li; Xu, Xiaoqin; Tao, Sijue; Xu, Fuqiang

doi:10.3389/fnana.2019.00063

Cited by 25 publications

(15 citation statements)

References 34 publications

(46 reference statements)

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“…Because PRV can infect neurons and cross synaptic connections in the nervous system, it has been widely used as a tracer for mapping neuronal circuits . PRV‐Bartha, one of the most commonly used viruses in neuronal tracing, is an attenuated vaccine strain that is only retrogradely spread along synaptic connections . As viruses can spread between cells either by diffusion in the extracellular space or by direct cell–cell transmission, many questions remain on how neuronal tracing viruses achieve a highly directional transport from cell to cell.…”

Section: Introductionmentioning

confidence: 99%

Proteomics Analysis Identifies IRSp53 and Fascin as Critical for PRV Egress and Direct Cell–Cell Transmission

Miao

Xia

et al. 2019

Proteomics

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Pseudorabies virus (PRV) has been widely used as a live trans‐synaptic tracer for mapping neuronal circuits. Systematically identifying mature PRV virion proteomes and defining co‐purified host proteins are necessary to fully understand the detailed mechanism underlying PRV transmission processes. Here, a PRV virion purification strategy based on sorting with flow cytometry is developed and the mature extracellular and intracellular PRV virion proteomes using LC coupled with MS/MS are characterized. In addition to viral proteins, a large number of host proteins are also identified, including proteins related to actin cytoskeletal dynamics and membrane protrusion. How many of these host proteins are true virion components are unknown and the majority of these may not be. Through functional analysis, it is found that IRSp53 and fascin are critical for the egress process and play a role in direct cell–cell transmission. Moreover, it is shown that CDC42 and Rac1 are also involved in the production of mature extracellular virions. The results suggest that the formation of the filopodia‐like cytoskeleton and the rearrangement of the membrane, which are both associated with IRSp53 and fascin, may be important for the transmission of viruses used in neuronal tracing.

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Section: Introductionmentioning

confidence: 99%

Proteomics Analysis Identifies IRSp53 and Fascin as Critical for PRV Egress and Direct Cell–Cell Transmission

Miao

Xia

et al. 2019

Proteomics

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“…In rats and mice, depending on the inoculation route, PRV infects the neuronal region, which receives fibers from or project to the site of inoculation. Thus, rats and mice are used as standardized animal models either to study alphaherpesviral neuroinvasion, the host’s response towards infection [ 24 , 25 , 140 ], or to analyze and identify neuroanatomical networks [ 141 ]. PRV mouse models can also be helpful to provide insights into the mechanisms underlying human alphaherpesvirus induced diseases such as HSV-1 encephalitis [ 27 ] or neuropathies after infection with the closely related varicella zoster virus [ 142 ].…”

Section: Discussionmentioning

confidence: 99%

Comparative Pathology of Pseudorabies in Different Naturally and Experimentally Infected Species—A Review

Sehl

Teifke

2020

Pathogens

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The pseudorabies virus (PRV) is an alphaherpesvirus and the causative agent of Aujeszky’s disease (AD). PRV infects a wide range of animal species including swine as the natural host as well as ruminants, carnivores, rodents and lagomorphs. In these species, except for the pig, PRV infection causes acute, severe disease, characterized by insatiable itching, and is always lethal. Horses, chickens and non-human primates have been shown to be largely resistant to PRV infection, while disease in humans is still controversial. PRV is a pantropic virus, which preferably invades neural tissue, but also infects epithelia of various organs, whereupon multisystemic lesions may result. Although AD is mainly associated with severe pruritus, also known as “mad itch”, there are notable differences regarding infection route, clinical signs, viral distribution and lesion patterns in different animal species. In this comprehensive review, we will present clinico-pathologic findings from different species, which have been either shown to be susceptible to PRV infection or have been tested experimentally.

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“…Mapping brain connectome requires both retrograde and anterograde trans-synaptic neuronal circuit tracers [3,12,13]. In retrograde trans-synaptic labeling system, pseudorabies virus (PRV) as a trans-multi-synaptic tracer, is widely used in mapping the connection between peripheral-central and central connections [14,15]. Rabies virus (RV), a rigorous transmono-synaptic tracer, is broadly used to analyze input brain circuits [4].…”

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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Optimization of the Fluorescent Protein Expression Level Based on Pseudorabies Virus Bartha Strain for Neural Circuit Tracing

Cited by 25 publications

References 34 publications

Proteomics Analysis Identifies IRSp53 and Fascin as Critical for PRV Egress and Direct Cell–Cell Transmission

Proteomics Analysis Identifies IRSp53 and Fascin as Critical for PRV Egress and Direct Cell–Cell Transmission

Comparative Pathology of Pseudorabies in Different Naturally and Experimentally Infected Species—A Review

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

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