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. 2019 May 16;216(7):1542–1560. doi: 10.1084/jem.20182386

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© 2019 Dodiya et al.

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Figure 2. — Reductions in Aβ pathology and altered plaque-localized microglial phenotypes are only observed in ABX-treated male APPPS1-21 mice. (A) Representative images of Aβ plaque burden in the cortex of vehicle (a and c) and ABX-treated (b and d) in 7-wk-old APPPS1-21 mice using anti-Aβ monoclonal antibody, 3D6. (a and b) Male vehicle vs. ABX, respectively; (c and d) female vehicle vs. ABX, respectively. (e) Quantification of plaque burden in vehicle- and ABX-treated APPPS1-21 mice using threshold-limited particle analysis of 3D6⁺-positive staining from six sections per case and expressed relative to the total cortical area of each slice. Aβ burden analysis revealed a significant effect of ABX (two-way ANOVA: F_{[1, 36]} = 10.58, P = 0.0025) and an interaction (F_{[1, 36]} = 9.145, P = 0.0046) while sex showed no significant effect (F_{[1, 36]} = 1.76, P = 1929). (B) Representative images of microglial cells using Iba1 antibody. No differences in total Iba1-positive cells were observed in vehicle versus ABX treatment irrespective of sex (male: a, b, and e; female: c, d, and e; two-way ANOVA—ABX treatment: F_{[1, 36]} = 0.036, P = 0.85; or sex: F_[1,36] = 0.19, P = 0.668; or an interaction between ABX treatment and sex: F_[1,36] = 0.131, P = 0.718). In contrast, the area of Iba1-positive cell bodies was significantly reduced in ABX treated male mice (b and f) but not in females (d and f; two-way ANOVA—ABX treatment: F_{[1, 36]} = 14.77, P = 0.0005; sex: F_{[1, 36]} = 4.97, P = 0.0321; and an interaction: F_{[1, 36]} = 142.2, P < 0.0001). Control male mice showed larger cell bodies compared with control female mice (P < 0.0001). ABX treatment resulted in significantly smaller cell bodies in males (P < 0.0001) and larger cell bodies in females (P < 0.0001) compared with their respective controls. (C) Imaris 3D reconstruction showed significantly different microglia morphology characteristics in males (a and b) compared with females (c and d) after ABX treatment (microglial branch length—two-way ANOVA: F_{[1, 36]} = 70.77, P < 0.0001; microglial branch end points: F_{[1, 36]} = 61.24, P < 0.0001), with sex (microglial branch length: F_{[1, 36]} = 91.42, P < 0.0001; microglial branch end points: F_{[1, 36]} = 75.38, P < 0.0001), and an interaction (microglial branch length: F_{[1, 36]} = 29.63, P < 0.0001; microglial branch end points: F_{[1, 36]} = 22.73, P < 0.0001). Vehicle-treated male mice showed significantly longer branch lengths (a; quantified in e, P = 0.029) and number of branch points (a; quantified in f, P = 0.04) compared with vehicle-treated females (c). ABX treatment resulted in significantly longer branch lengths (b; quantified in e, P < 0.0001) and increased numbers of branch points (b; quantified in f, P < 0.0001) in male mice. Scale bars in A (a–d), B (a–d), and C (a–d) represent 500 µm, 10 µm, and 10 µm, respectively. Data are mean ± SEM. *, P < 0.05; ***, P < 0.001; ****, P < 0.0001; n = 10/group.