Technical Reference #1713

1713.

Virion-Incorporated Glycoprotein B Mediates Transneuronal Spread of Pseudorabies Virus Dusica Curanovic and Lynn W. Enquist, Princeton University, Journal of Virology, 83(1713), (2009)
Link To Paper

Abstract:
Transneuronal spread of pseudorabies virus (PRV) is a multistep process that requires several virally encoded proteins. Previous studies have shown that PRV glycoprotein B (gB) a component of the viral fusion machinery is required for the transmission of infection to postsynaptic second-order neurons. We sought to identify the gB-mediated step in viral transmission. We determined that gB is not required for the sorting of virions into axons of infected neurons anterograde transport or the release of virions from the axon. trans or cis expression of gB on the cell surface was not sufficient for transneuronal spread of the virus; instead efficient incorporation of gB into virions was required. Additionally neuron-to-cell spread of PRV most likely does not proceed through syncytial connections. We conclude that upon gB-independent release of virions at the site of neuron-cell contacts the virion-incorporated gB/gH/gL fusion complex mediates entry into the axonally contacted cell by fusion of the closely apposed membranes.

Materials & Methods:
Live-cell imaging. Dissociated SCG neurons were plated onto glass-bottom MatTek dishes and infected at a high multiplicity of infection for 16 h. Live-cell imaging was performed as described previously (10). Imaging was performed on a Perkin-Elmer RS3 spinning-disk confocal microscope with a 60 oil objective (numerical aperture 1.3).

Microscopic Technique
Fluorescence Microscopy, Immunofluorescence, Live Cell Imaging

Cell Type(s)
Dissociated SCG neurons