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. 2021 Feb 19;13:593927. doi: 10.3389/fnagi.2021.593927

FIGURE 7.

FIGURE 7

Alzheimer’s disease EVs but not control EVs injection cause GS astrocyte alteration and higher vessel diameter in wild-type mouse brains. (A) A schema illustrating EVs from human A-EVs bilaterally injected to the hippocampus of C57BL6 mice at 10 months of age. (B) Representative of maximal Z-projection of Hoescht (blue), CLN-5 (green), GS (red) staining 48 h after intrahippocampal injection of CNT, FAD- and SAD A-EVs into aged C57BL6 mouse brain. Original magnification: × 60 (zoom 1.6). White arrowheads indicate regions where high vascular diameter, while empty arrowheads point to increased GS. (C) Quantification of the Z projected intensity of GS. ANOVA comparations of A-EVs FAD with ∗ A-EVs Control and # A-EVs SAD, levels of significance were set to *p < 0.05, **p < 0.01, and ###p < 0.001. (D) Quantification of vessels diameter using of the CLN-5 Z projected images. ∗Comparison with AEVs Control, levels of significance were set to **p < 0.01. (E) Segmented images of GS endfeet (white) contacting CLN-5 vessel (blue). White arrowheads indicate regions where focal GS vascular coincides with CLN-5 staining. The insets are crop magnifications of double positive GFAP-CLN-5 areas (white). (F) Quantification of the endfeet volume of double positive GFAP-CLN-5. ∗Comparison with A-EVs Control, levels of significance were set to *p < 0.05. Scale bar: (B,E), 20 μm. Data are presented as mean ± SEM from mice treated with CNT- (n = 3), SAD- (n = 3), and FAD- (n = 3) A-EVs; independent experiments.