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Figure 4 Microglial activation and neuroinflammation in the larval and adult sgshΔex5−6 CNS. (A,B) L-plastin immunostaining of microglia in wild type and sgshΔex5−6 7 dpf larval brains. Scale bar = 50 µm. (A`) Zoom of boxed region in (A) showing a representative ramified microglial cell in the larval zebrafish brain. (B`) Zoom of boxed region in (B) showing representative amoeboid microglial cell in the larval zebrafish brain. (C) Quantification of L-plastin+ microglial morphology and abundance in wild type or sgshΔex5−6 7 dpf brains. Each point represents a single brain. Data presented as mean ± SEM and tested by ordinary one-way ANOVA with Tukey’s multiple comparisons test; **** p < 0.0001. (D–E) L-plastin immunostaining of microglia in midbrain sections of adult wild type and sgshΔex5−6 zebrafish, scale bar = 50 µm. (D`) Zoom of boxed region in (D) showing representative ramified microglial cell in the adult zebrafish midbrain. (E`) Zoom of boxed region in (E) showing representative amoeboid microglial cell in the adult zebrafish midbrain. (F) Quantification of L-plastin+ microglial morphology and abundance in adult wild type and sgshΔex5−6 midbrain sections. Each point represents a single section, n = 3 sections of approximately equal level quantified per brain. Data presented as mean ± SEM and tested by ordinary one-way ANOVA with Tukey’s multiple comparisons test; **** p < 0.0001. (G–I) Detection by TEM of perivascular microglia with activated morphology with large, electron-lucent intracellular vacuoles (arrows) in the midbrain of 18-month-old sgshΔex5−6 zebrafish. Scale bars 2 µm in (G), 100 µm in (H,I).