Pseudorabies Virus Infection Alters Neuronal Activity and Connectivity In Vitro
Figure 5
Increasingly larger molecular weight dyes transfer between neurons as infection proceeds.
(A) Lucifer yellow (LY, 457 MW) dye did not transfer from filled mock-infected cell bodies to other cell bodies (0/10). Images represent overlays of excited Lucifer yellow onto bright field images. (B–C) Transfer of LY to other cell bodies could be seen in PRV 151 infected neurons by 9 hpi,(2/12). White arrows indicate cell bodies that contain dye transferred from the filled neuron. Transfer of LY from filled cell bodies to other cell bodies was more common in PRV 151 infected neurons by 24 hpi (2/6). Note that PRV 151 infected cell bodies that contained transferred dye were generally not adjacent to filled cell body, and were exclusively to non-adjacent cell bodies early (9–12 hpi). (D) Percentage of neurons assayed in which dye spread from the filled cell body to another cell body. (E) Texas-red dextran conjugates (3,000–40,000 MW) to not transfer from filled cell bodies of mock-infected neurons to other cell bodies (0/9). (F–H) Large dye transfer between infected neurons occurs late in infection (14–16 hpi, 0/9, 18–20 hpi, 0/9, 24–26 hpi, 11/19). Neurons selected for dye filling were chosen as potentially fused cells based on outer membrane contours by light microscopy. Green – GFP expressed from viral genome, Red – Texas-red dextran conjugate, Yellow – overlap of green and red images. Panel F, bright field image was recorded following fluorescence image and pipette removal; pipette location is indicated by a dashed while line.