Figure 4.

Zebrafish: Early microglia phenotype. (A) Brightfield images of wt and itgav^−/− zebrafish at 5 dpf demonstrating intracranial hemorrhage, swollen hearts, and gut abnormalities in the knockouts (arrowheads). (B) Volcano plots of differentially expressed transcripts in 8 dpf itgav^−/− compared with wt whole zebrafish, annotated on GRCz11 (left). (n = 3, adjusted P value 0.05, DESeq2 Wald). A subset of genes related to TGF-β signaling (right). (C) Heatmap of the top 30 unbiased differentially expressed genes in 8 dpf itgav^−/− larvae (n = 3). (D–F) Representative images of the eye vessels of itgav^−/−;Tg(flk1:EGFP) and wt;Tg(flk1:EGFP) zebrafish larvae at 6 dpf. Confocal images of intraocular vasculature network (lateral view) of the whole mount. The intensity of EGFP of the intraocular vasculature (E) and the number of vessels branching in the intraocular vasculature network were quantified (F) (n = 4 zebrafish per group, t test, P < 0.01, P < 0.05, error bars mean ± SEM). (G and H) Representative images of zebrafish larvae microglia (arrowheads) stained with neutral red. Dorsal view (right) and lateral view (left) of wt and itgav^−/− zebrafish microglia at 6 dpf. Quantification of microglia numbers is shown on the right. (n = 16, unpaired t test, P < 0.0001, mean).