Fig. 2: DBN controls astrocyte scar formation and astrocyte reactivity in vivo. | Nature Communications

Fig. 2: DBN controls astrocyte scar formation and astrocyte reactivity in vivo.

From: Drebrin controls scar formation and astrocyte reactivity upon traumatic brain injury by regulating membrane trafficking

Fig. 2

A GFAP+ astrocytes at core lesion sites in WT (upper panels) and Dbn−/− (lower panels) brains, 7 days post stab injury (DPI). Magenta lines indicate the stab injury from needle insertion. Magnifications show stab lesions, with “palisading” astrocytes extending towards injury sites in WT mice. Scale bars: 100 µm. Quantification of palisading astrocytes in stab wounds 7 DPI, bar graph shows mean, individual data points and SEM. n = 4 WT and 3 Dbn−/− animals, Student’s unpaired t test, two-sided, **P = 0.0016 t = 6.205 df = 5). B IBA1 + microglia (magenta) and GFAP+ reactive astrocytes (green) in the periphery of scars 7 DPI, in WT (upper panel) and Dbn−/− brains (lower panel). Scale bars: 10 µm. Quantification of soma sizes of IBA1+ microglia. n = 145 WT & 122 Dbn−/− cells from three mice per condition, graph shows box and whisker plots: box extends from 25th to 75th percentiles, central line = median, whiskers comprise all values from minimum to maximum, F test ***< 0.0001, Unpaired t test with Welch’s correction, two-sided; ***P = 0.000000205637175, t = 5.381 df = 201). C NeuN+ neurons (magenta) and GFAP+ reactive astrocytes (green) in scars of WT (upper panel) and Dbn−/− brains (lower panel) 7 DPI. Scale bars: 10 µm. Quantification shows neurons without nuclear NeuN in scars. n = 3 WT and three Dbn−/− mice, bar graph shows mean, individual data points and SEM, Unpaired t test with Welch’s correction, two-sided; *P = 0.0231, t = 6291 df = 2044). D IHC of WT and DBN−/− brains with 30-day post stab lesions. GFAP labeling (green) detects reactive astrocytes; SOX9 labeling (magenta) detects astrocytes. Scale bars in overview images: 200 µm. Scale bars in close up images: 10 µm. Quantification of cytoplasmic/nuclear SOX9 distribution in astrocytes in lesion sites. Single data points of nuclear SOX9 and corresponding error bars are displayed in magenta, cytoplasmic SOX9, and corresponding error bars in green. n = 4–5 animals per condition. E NeuN+ neurons (magenta) and GFAP+ reactive astrocytes (green) in WT (left), Dbn−/− (center) and Dbnfl/fl:CAMK-Cre+/cre brains (right), 30 DPI. Quantifications show the percentage of animals with GFAP+ astrocytes (left graph) and NeuN+ cells (right graph) at lesion sites 30 days post stab wounding. Bar graph shows mean, individual data points and SEM n = 4 WT, five Dbn−/− animals, six Dbnfl/fl:CAMK-Cre animals, (one-way ANOVA DFn = 2, DFd = 12, F = 8.397; Dunnett’s multiple comparisons test Multiplicity adjusted p value: *P = 0.0112). All images in this figure are confocal stacks. Scale bars: 200 µm. Source data are provided as a Source Data file.

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