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. 2023 Jul 10;120(29):e2302892120. doi: 10.1073/pnas.2302892120

Fig. 2.

Fig. 2.

Sex-specific differences in microglial mobility after focal vascular injury. (A) In vivo two-photon images showing a microglia (yellow arrow) in a male mouse migrate toward a microbleed over a 24-h period. Note that vessel ablation leads to red-emitting autofluorescence at the injury site (yellow asterisks). (Scale bar, 20 µm.) (B) Fraction of mobile and stationary microglia imaged over a 24-h period under normal (no bleed; 272 cells from nine mice) or Bleed (267 cells from 13 mice) conditions, with data from both sexes pooled together. Chi-square analysis indicates the fraction of mobile microglia increased significantly after injury (χ2 = 109.9, P < 0.01). (C) Bar graphs show the absolute distance traveled by mobile microglia in No Bleed (15 cells) and Bleed (54 cells) conditions for each 12-h interval or the sum over 24 h, with data from both sexes pooled together. Data are mean ± SEM. (D) The net distance traveled by mobile microglia toward or away from the injury site (or an equivalent landmark in controls), with data from both sexes pooled together. Data are mean ± SEM. (E) Graphs show the absolute movement of each mobile microglia plotted over time (E, Left) or the group average (E, Right) after injury, stratified by sex (32 cells from seven males and 22 cells from six female mice). Data are mean ± SEM. (F) The net movement of each mobile microglia plotted over time (F, Left) or the group average (F, Right) after injury, stratified by sex. Data are mean ± SEM. (G) Fraction of mobile or stationary microglia for male and female mice after injury. Chi-square analysis indicates males had more mobile microglia after injury than female mice (χ2 = 9.33, P < 0.01). (H) Linear regression analysis in both male and female mice shows the relationship between how far a microglia moves over a 24-h period as a function of each cell’s initial distance from the bleed. Note the moderate and significant relationship in male but not female mice. Large open circles represent mobile microglia while smaller dots with dark outline represent stationary microglia. Statistical analysis for C–F was conducted with two-tailed unpaired t tests.