Anterograde or retrograde transsynaptic labeling of CNS neurons with vesicular stomatitis virus vectors

Abstract

To understand how the nervous system processes information, a map of the connections among neurons would be of great benefit. Here we describe the use of vesicular stomatitis virus (VSV) for tracing neuronal connections in vivo. We made VSV vectors that used glycoprotein (G) genes from several other viruses. The G protein from lymphocytic choriomeningitis virus endowed VSV with the ability to spread transsynaptically, specifically in an anterograde direction, whereas the rabies virus glycoprotein gave a specifically retrograde transsynaptic pattern. The use of an avian G protein fusion allowed specific targeting of cells expressing an avian receptor, which allowed a demonstration of monosynaptic anterograde tracing from defined cells. Synaptic connectivity of pairs of virally labeled cells was demonstrated by using slice cultures and electrophysiology. In vivo infections of several areas in the mouse brain led to the predicted patterns of spread for anterograde or retrograde tracers.

The brain parenchyma has a type I interferon response that can limit virus spread

“…The infection and spread of replication-competent rVSV-eGFP was quantified. The CP was chosen as an inoculation site due to its unidirectional, anterograde connections to the globus pallidus (GP) and basal nuclei (NB) (15).…”

“…We were able to evaluate the effectiveness of the antiviral response mounted within the brain parenchyma by exploiting the characteristics of a transsynaptic virus and the well-defined connections of the basal ganglia (15). This approach enabled the histological identification and quantification of the types of cells that were infected, which included neurons infected by transsynaptic spread.…”

“…VSV is the prototypic nonsegmented negative-strand RNA virus and has been used extensively to study the systemic innate immune response, which promotes its rapid clearance (13,14). VSV was selected because it has a characteristic transsynaptic anterograde transmission pattern among neurons and is straightforward to track, as it can be engineered to express eGFP (15). As the spread of wildtype (WT) VSV is not well-controlled in the mouse brain and can rapidly lead to mortality (16), we sought to provide the host with a "head start" in the induction of an innate immune response following infection.…”

Spontaneous cortical activity alternates between motifs defined by regional axonal projections

“…With the use of retrograde polysynaptic rabies virus tracers 42 , it would be possible to target multi-synaptic pathways. These experiments could also involve rabies viruses that permit both structural and functional analysis using optogenetic tools to manipulate upstream projections 43 .…”

Viruses in human central nervous system — invasion, transmission, and latency: a literature review

“…У многих покрытых вирусов -RABV, MV окончательная сборка и выход вируса из клетки -координирован-ный процесс. Вирусные компоненты -нуклеокапсид и мембранные белки транспортируются отдельно, концентрируются в дендрите или аксоне, собираются в клеточной мембране, после чего вирус выходит из клетки [49,50].…”