Low MOI axonal PRV infection in compartmented neuronal cultures results in a quiescent infection in a small number of neurons.

<p><b>(A)</b> Tri-chamber compartments, S: soma (cell bodies), M: methocel (middle), N: axonal. PRV 233 infection was made in the N compartment at an MOI of 0.01. DiI (red) was added to the N compartment media 6 hpi to label red cell bodies that project axons to the N compartment. Circles highlight primarily infected neurons. Insets show single infected neurons (green) among surrounding non-infected neurons. <b>(B)</b> GFP positive, DiI labeled or non-labeled cell bodies were counted at 3 dpi (separate or merged channels are shown, arrowhead points to one GFP positive cell body, ph: phase contrast). Ratios of either red cell bodies to all S compartment neurons (connectivity), or green cell bodies to dual color (green and red) cell bodies (infectivity) were calculated and shown in the graph. <b>(C)</b> RNA was isolated after 7 days of either S compartment or N compartment infection with PRV180 at an MOI of 0.01. LAT and EP0 transcripts were quantitated. S compartment (cell body) infection resulted in a productive infection (red capsid accumulation in all of the cells), whereas N compartment (axonal) infection was silent (no detectable red fluorescent signal anywhere in the S compartment). The graph shows the ratio of each transcript in axonal to cell body infection after normalization to 28S rRNA. <b>(D)</b> Illustration of the complementation assay: N compartments are infected with PRV180 at an MOI of 0.01 while S compartments are treated with drugs, inhibitors, viruses or virus-like particles.