Characterization of the neuroinvasive profile of a pseudorabies virus recombinant expressing the mTurquoise2 reporter in single and multiple injection experiments.

TitleCharacterization of the neuroinvasive profile of a pseudorabies virus recombinant expressing the mTurquoise2 reporter in single and multiple injection experiments.
Publication TypeJournal Article
Year of Publication2018
AuthorsHogue, IB, J Card, P, Rinaman, L, Goraczniak, HStaniszews, Enquist, LW
JournalJ Neurosci Methods
Volume308
Pagination228-239
Date Published2018 10 01
ISSN1872-678X
KeywordsAnimals, Brain, Cell Line, Genetic Vectors, Herpesvirus 1, Suid, Male, Neural Pathways, Neuroanatomical Tract-Tracing Techniques, Neuronal Tract-Tracers, Neurons, Rats, Sprague-Dawley, Viscera
Abstract

<p><b>BACKGROUND: </b>Viral transneuronal tracing has become a well established technology used to define the synaptic architecture of polysynaptic neural networks.</p><p><b>NEW METHOD: </b>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.</p><p><b>CONCLUSION: </b>Collectively, these data demonstrate that PRV-290 is a valuable addition to the viral tracer toolbox for transneuronal tracing of neural circuitry.</p>

DOI10.1016/j.jneumeth.2018.08.004
Alternate JournalJ. Neurosci. Methods
PubMed ID30098326
PubMed Central IDPMC6294127
Grant ListR01 NS033506 / NS / NINDS NIH HHS / United States
R01 DK100685 / DK / NIDDK NIH HHS / United States
R01 MH059911 / MH / NIMH NIH HHS / United States
R01 NS060699 / NS / NINDS NIH HHS / United States
P40 RR018604 / RR / NCRR NIH HHS / United States
P40 OD010996 / OD / NIH HHS / United States