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Fig. 2 | Acta Neuropathologica Communications

Fig. 2

From: Regulating microglial miR-155 transcriptional phenotype alleviates Alzheimer’s-induced retinal vasculopathy by limiting Clec7a/Galectin-3+ neurodegenerative microglia

Fig. 2

Conditional depletion of microglial miR-155 decreases Galectin-3+ microglia and upregulates homeostatic P2ry12 expression in APP/PS1 mice retinas. ae Representative images of immunofluorescent staining for Galectin-3+ microglia (red), Tmem119+ microglia (green) and lectin for blood vessels (blue) on retinal flat mounts from a APP/PS1:miR-155cKO genotype, inner plexiform layer (IPL); b APP/PS1 genotype, ganglion cell layer (GCL) to IPL; c APP/PS1 genotype, outer plexiform layer (OPL); d APP/PS1:miR-155cKO genotype, IPL; and e APP/PS1 genotype, IPL. Images were obtained using 20 × or 63 × microscope objectives. Dashed-line rectangles highlight Galectin-3+ microglia. Scale bars = 10 µm. f Quantitative analysis of 12F4 for Aβ42 immunoreactivity (IR) in retinal cross-sections of mice from all experimental groups (n = 48 total, n = 6 each group). g Manual counting of Galectin-3+ microglia in each retinal cross-section from all experimental groups of the same mouse cohort. h Representative images of immunofluorescent staining for Galectin-3+ microglia (red), Iba1+ microglia (green), 12F4 for Aβ42 (white) and DAPI (blue) on retinal cross-sections from APP/PS1:miR-155cKO and APP/PS1 mice. Images were obtained using 40 × microscope objectives. Dashed-line rectangle highlights a Galectin-3+ microglia engulfing Aβ42. Scale bars = 10 µm. i, j. Representative images of eye cross-section depicting immunofluorescent staining for i Tmem119+ microglia (red), 12F4 for Aβ42 (green) and DAPI (blue) from an APP/PS1:miR-155cKO mouse and j P2ry12+ microglia (red), 12F4 for Aβ42 (green) and DAPI (blue) from an APP/PS1 mouse. k Manual counting of Tmem119+ microglia in each retinal cross-section from all experimental groups of the same mouse cohort. l Densitometric analysis of western blotting protein bands of P2ry12 normalized by β-actin control for retinal lysates from all experimental groups (n = 48 total, n = 6 each group). m Manual counting of Apoe+ microglia in each retinal cross-section from all experimental groups of the same mouse cohort shown in figures g and k n Densitometric analysis of western blotting protein bands of Iba1 normalized by β-actin control for retinal lysates from all experimental groups of the same cohort as figure l. Data from individual mice (circles) as well as group means ± SEMs are shown. Black-filled circles represent male and clear circles represent female animals. *p < 0.05, **p < 0.01, by two-way or three-way ANOVA with Tukey’s post-hoc multiple comparison test. Two group statistical analysis was performed using an unpaired two-tailed Student t-test and is shown in parentheses. Fold changes and percentage decreases are shown in red

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