FIG 6.
Laser scanning confocal microscopy for PrPSc and LAMP-1 in nerve bundles of the tongue. Laser scanning confocal microscopy for LAMP-1 (A, D, and G), PrPSc (B, E, and H), and both LAMP-1 and PrPSc (C, F, and I) in nerve bundles in tongue from HY TME-infected hamsters was performed. Panels A through C, D through F, and G through I are the same fields of view within their respective groups. In nerve bundles, LAMP-1 immunofluorescence was observed in a discrete pattern within the nerve bundle and in association with axons. A cluster of LAMP-1 immunofluorescence was also observed in areas outside axons that were consistent with localization to the cell body of Schwann cells based on the proximity of immunofluorescence to nuclei (stained with ToPro-3 and shown in blue). A white arrow within a panel series indicates the area of colocalization of PrPSc and LAMP-1. Colocalization analysis on 27 separate three-dimensional (3D) reconstructions for PrPSc and LAMP-1 revealed that there was an overlap of PrPSc with LAMP-1 in nerve bundles (Table 3). Panels A to C represent a single image that was taken from a 3D reconstruction of an LSCM stack assembled from 64 individual images. This 3D reconstruction of nerve bundles stained for PrPSc and LAMP-1 is illustrated in Movie S4 in the supplemental material. Scale bars, 5 μm (A and D) and 1 μm (G).