Fig. 3
Retinal amyloidosis in blood vessel walls of APPSWE/PS1ΔE9 (ADtg) mice. a–b Representative fluorescence images acquired using a 20 × or b 63 × microscope objectives, of isolated retinal microvasculature stained for Aβ (4G8, magenta), blood vessels (lectin, green), and nuclei (DAPI, blue) in age- and sex-matched perfused ADtg mice (n = 6) and wild-type littermates (WT; n = 6). Scale bars = 20 µm. c–d Representative fluorescence virtual cross-section images acquired using the Leica confocal microscope 63 × objective of the isolated retinal microvasculature stained for Aβ (4G8, magenta), blood vessels (lectin, green) and nuclei (DAPI, blue) in an 8-month-old male ADtg mouse. Scale bar = 20 µm. Arrows indicate vascular Aβ. e Quantitative analysis of Aβ (4G8)-immunoreactive (IR) area in each microscopic field of isolated retinal microvasculature from WT vs. ADtg mice. f Quantitative analysis of the Aβ (4G8)-IR area stratified by mice age group (8 months vs. 12 months) and genotype (WT vs. ADtg) in the same cohort. g–g′. Representative fluorescence images of the isolated retinal microvasculature stained for Aβ (4G8, red), blood vessels (lectin, green) and nuclei (DAPI, blue) in an 8-month-old male ADtg mouse; Aβ signals occur in retinal vessel walls and vascular cells. h–i. Representative fluorescence images of fixed retinal cross-section stained for thioflavin-S (Thio-S, green), Aβ40 (11A50-B10, red) and blood vessels (CD31, blue) in an 8-month-old male ADtg mouse showing h vertical blood vessel and i longitudinal blood vessel. Data from individual mice (circles) as well as group means ± SEMs are shown. Black-filled circles represent males and clear circles represent females. Fold changes are shown in red. *p < 0.05, **p < 0.01, by two-way ANOVA with Tukey’s post hoc multiple comparison test or by an unpaired 2-tailed Student t-test. P and F values refer to comparisons of age groups (PA, FA), ADtg versus WT genotype groups (PG, FG), and interactions (PI, FI)