Genetically timed, activity-sensor and rainbow transsynaptic viral tools

Abstract

We developed retrograde, transsynaptic pseudorabies viruses (PRVs) with genetically encoded activity sensors that optically report the activity of connected neurons among spatially intermingled neurons in the brain. Next we engineered PRVs to express two differentially colored fluorescent proteins in a time-shifted manner to define a time period early after infection to investigate neural activity. Finally we used multiple-colored PRVs to differentiate and dissect the complex architecture of brain regions.

Microdissection of neural networks by conditional reporter expression from a Brainbow herpesvirus

“…The ability of neurotropic viruses to replicate and spread through neural circuits has brought a polysynaptic perspective to circuit analysis (2-6). Recent genetic engineering of such viruses has produced increasingly powerful probes that have provided novel insights into the identity, organization, and activity of neural networks in a variety of systems (7)(8)(9).…”

Studies on the role of disturbed Ca2+ homeostasis in the pathomechanism of the cardiac effects of experimental diabetes using conventional and novel experimental techniques”

“…Several previous reports have been successful in construction of PRVs for delivering foreign genes to neurons [35]. PRV is an especially important tool for labelling neuronal circuits [36], which was combined with delivery of activity markers to labelled neurons [37]. Application of PRV-based gene transfer vectors in the cardiovascular research field has not yet been reported.…”

Effect of us1 gene mutation on pseudorabies virus gene expression

“…Because of these drawbacks it is not recommended to use alpha-herpesviruses in higher doses and for longer time period [56]. Earlier, another problem was the bidirectional spreading, but today certain modified strains has the advantage of unidirectional transfer [50,57].In contrast, rabies does not change neuronal metabolism,…”