FIGURE 2.

Ex vivo WT and Tg mouse wholemount and 6E10 immunofluorescence. Aβ deposits in the ex vivo retinal wholemount were identified by 6E10 immunofluorescence (red) and confocal imaging taken at the level of the GCL. The confocal imaging was initiated at the NFL, and as the optical steps proceeded deeper into the wholemount, DAPI (blue) nuclear staining was used to identify the beginning, middle and end of the GCL. All images in Figure 2 are shown at the level of the mid GCL. (A) Representative confocal image after Aβ immunofluorescence of a younger (6 month) WT retina. Note the small specks of Aβ deposits in red immunofluorescence, some shown by white arrowheads. A larger opaque red profile is an artifact (white asterisk). (B) Representative image of a negative control section, in which the primary antibody was replaced with a non-specific IgG. Image demonstrates very low background levels of red immunofluorescence. (C) Representative image of Aβ immunoreactivity in the 6 month Tg retina with representative red specks of immunofluorescence shown by white arrowheads. (D) In an older (18 month) Tg retina, the retinal immunofluorescence increased with more red immunoreactive specks (white arrowheads) and an occasional immunoreactive retinal ganglion cell (yellow arrowhead). (A–D) Scale bar: 20 μm. (E) An orthogonal view of the confocal z-stack reconstruction from a 15 month Tg retina demonstrates that the majority of the Aβ immunoreactivity is present in the inner retina, specifically in the NFL and GCL. Some Aβ immunoreactive specks can be seen extending deeper into the IPL. Cell nuclear labeling with DAPI (blue) reveals lamination in the retina. Lamination and scale bars are shown on the right axes.