Characterization of the neuroinvasive profile of a pseudorabies virus recombinant expressing the mTurquoise2 reporter in single and multiple injection experiments. Author Ian Hogue, J Patrick Card, Linda Rinaman, Halina Goraczniak, Lynn Enquist Publication Year 2018 Type Journal Article Abstract BACKGROUND: Viral transneuronal tracing has become a well established technology used to define the synaptic architecture of polysynaptic neural networks.NEW METHOD: In this report we define the neuroinvasive profile and reporter expression of a new recombinant of the Bartha strain of pseudorabies virus (PRV). The new recombinant, PRV-290, expresses the mTurquoise2 fluorophor and is designed to complement other isogenic recombinants of Bartha that express different reporters of infection. Results & Comparison with Existing Methods: PRV-290 was injected either alone or in combination with isogenic recombinants of PRV that express enhanced green fluorescent protein (EGFP; PRV-152) or monomeric red fluorescent protein (mRFP; PRV-614). Circuits previously defined using PRV-152 and PRV-614 were used for the analysis. The data demonstrate that PRV-290 is a retrograde transneuronal tracer with temporal kinetics similar to those of its isogenic recombinants. Stable expression of the diffusible mTurquoise2 reporter filled infected neurons, with the extent and intensity of labeling increasing with advancing post inoculation survival. In multiple injection experiments, PRV-290 established productive infections in neurons also replicating PRV-152 and/or PRV-614. This novel demonstration of three recombinants infecting individual neurons represents an important advance in the technology.CONCLUSION: Collectively, these data demonstrate that PRV-290 is a valuable addition to the viral tracer toolbox for transneuronal tracing of neural circuitry. Keywords Animals, Cell Line, Male, Neural Pathways, Brain, Neurons, Rats, Sprague-Dawley, Herpesvirus 1, Suid, Genetic Vectors, Neuroanatomical Tract-Tracing Techniques, Neuronal Tract-Tracers, Viscera Journal J Neurosci Methods Volume 308 Pages 228-239 Date Published 2018 Oct 01 ISSN Number 1872-678X DOI 10.1016/j.jneumeth.2018.08.004 Alternate Journal J Neurosci Methods PMCID PMC6294127 PMID 30098326 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML