FIG 7.
Laser scanning confocal microscopy for PrPSc and either LAMP-1 or cathepsin D in ganglia of the tongue. Laser scanning confocal microscopy for LAMP-1 (A and D), PrPSc (B and E), and both LAMP-1 and PrPSc (C, F, and M) or for cathepsin D (G and J), PrPSc (H and K), and both cathepsin D and PrPSc (I, L, and N) in ganglia of the tongue from HY TME-infected (A to L) and mock-infected (M and N) hamsters was performed. Panels A through C and G through I are the same fields of view within their respective groups. The boxed area in panel C is enlarged in panels D through F, and the boxed area in panel I is enlarged in panels J through L. Both LAMP-1 and cathepsin D immunofluorescence had a punctate pattern that was widespread within the somata of neurons. Nuclei (blue) were stained with ToPro-3. A white arrow within a panel series indicates the area of colocalization of PrPSc and the organelle marker. Each group of panels represents a single image that was taken from a three-dimensional (3D) reconstruction of an LSCM stack assembled from 64 individual images. Colocalization analysis on 27 separate 3D reconstructions for each PrPSc and cell marker combination revealed that there was an overlap of PrPSc with either LAMP-1 or cathepsin D in somata of ganglia (Table 3). 3D reconstructions of ganglia stained for PrPSc and the cellular markers in this figure are illustrated in Movies S6 and S7 in the supplemental material. Scale bar, 10 μm.