Aβ deposition in ocular blood vessels and optic nerve meningeal lymphatic vessels of 5×FAD mice. (A and D) Representative confocal images of retinal and optic nerve flatmounts from 10-mo-old WT and 5×FAD mice stained with 6E10 (red) for Aβ and IB4 (green) for blood vessels. (B and E) Statistical graphs showing the percentage of 6E10+ area in the retina and optic nerve (n = 10). (C) Fluorescence intensity profile through a line scan across retinal blood vessels, demonstrating co-localization of 6E10 and IB4. (F) 3D reconstruction of retina and optic nerve head (ONH) immunofluorescence pictures of 5×FAD mice labeled with 6E10 and IB4. White arrowheads indicate the majority of Aβ in the PVS. (G and H) ELISA of Aβ1–40 and Aβ1–42 in retinal samples from 10-mo-old WT mice and 5×FAD mice (n = 6). (I and J) Immunofluorescent staining of retina and optic nerve frozen sections from 5×FAD mice reveals Aβ1–42 (red) deposition at blood vessels, with αSMA (green)-positive and CD31 (blue)-positive labeling indicating arteries (A), and CD31 (blue) -positive and αSMA (green)-negative labeling indicating veins (V). (K) Fluorescence intensity line graph indicated that Aβ deposition in the PVS adjacent to arterioles was greater than that near venules, while deposition within the walls of venules was greater than inside the arterioles (n = 6–7). (L) Representative immunofluorescence images of 5×FAD mouse optic nerve tissue sections stained with 6E10 (red), MBP (purple), and TUJ1 (green). Aβ deposits in the spaces between myelinated axons were indicated by the arrowheads. (M and N) Representative immunofluorescence images showing Aβ labeled deposition with 6E10 (gray) in the optic nerve meningeal lymphatics and periorbital lymphatics in 5×FAD mice. The arrowheads indicated Aβ (gray) deposits colocalized with LYVE1+ (purple) and Laminin− (green) labeled lymphatic vessels. Data are representative of two independent experiments. Data are presented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t tests (B, E, G, and H). GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer.
Figure 2.

Aβ deposition in ocular blood vessels and optic nerve meningeal lymphatic vessels of 5×FAD mice. (A and D) Representative confocal images of retinal and optic nerve flatmounts from 10-mo-old WT and 5×FAD mice stained with 6E10 (red) for Aβ and IB4 (green) for blood vessels. (B and E) Statistical graphs showing the percentage of 6E10+ area in the retina and optic nerve (n = 10). (C) Fluorescence intensity profile through a line scan across retinal blood vessels, demonstrating co-localization of 6E10 and IB4. (F) 3D reconstruction of retina and optic nerve head (ONH) immunofluorescence pictures of 5×FAD mice labeled with 6E10 and IB4. White arrowheads indicate the majority of Aβ in the PVS. (G and H) ELISA of Aβ1–40 and Aβ1–42 in retinal samples from 10-mo-old WT mice and 5×FAD mice (n = 6). (I and J) Immunofluorescent staining of retina and optic nerve frozen sections from 5×FAD mice reveals Aβ1–42 (red) deposition at blood vessels, with αSMA (green)-positive and CD31 (blue)-positive labeling indicating arteries (A), and CD31 (blue) -positive and αSMA (green)-negative labeling indicating veins (V). (K) Fluorescence intensity line graph indicated that Aβ deposition in the PVS adjacent to arterioles was greater than that near venules, while deposition within the walls of venules was greater than inside the arterioles (n = 6–7). (L) Representative immunofluorescence images of 5×FAD mouse optic nerve tissue sections stained with 6E10 (red), MBP (purple), and TUJ1 (green). Aβ deposits in the spaces between myelinated axons were indicated by the arrowheads. (M and N) Representative immunofluorescence images showing Aβ labeled deposition with 6E10 (gray) in the optic nerve meningeal lymphatics and periorbital lymphatics in 5×FAD mice. The arrowheads indicated Aβ (gray) deposits colocalized with LYVE1+ (purple) and Laminin− (green) labeled lymphatic vessels. Data are representative of two independent experiments. Data are presented as mean ± SEM. Statistical analysis was performed using two-tailed unpaired t tests (B, E, G, and H). GCL, ganglion cell layer; IPL, inner plexiform layer; INL, inner nuclear layer; OPL, outer plexiform layer; ONL, outer nuclear layer.

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