Fig. 4. Retina hyaloid vascular alterations were enhanced by endogenous pdx1 expression silencing in aldh3a1−/− Tg(fli1:EGFP) zebrafish larvae. Endogenous pdx1 expression silencing by pdx1 morpholino injection led to widening IOC branch diameters and increased sprouts formation in hyaloid vasculature of aldh3a1−/− Tg(fli1:EGFP) zebrafish larvae at 120 hpf. (A). Representative confocal scans of the isolated hyaloid vasculature in aldh3a1+/+ and aldh3a1−/− larvae after Control-Mo or SB-pdx1-Mo injection. At 120 hpf, the hyaloid network has a basket-like structure branching off at the central hyaloid/optic artery (white box) and connects to the circumferential inner annular hyaloid vessel (white arrow) encompassing the lens, also regularly called IOC in the matured adult retinal vasculature. Several sprouts (white delta) interconnect the basket-like vascular arcades and indicate the angiogenesis directly. White scale bar = 20 μm. (B–C). Quantification of the IOC branch diameter and sprouts formation in violin plots, n = 13–15 per group. 6 ng of morpholinos: Control-Mo and SB-pdx1-Mo were injected into the one-cell stage of zebrafish embryos, respectively. For statistical analysis one-way ANOVA followed by Sidak's multiple comparison test was applied, **p < 0.01, ***p < 0.001, ****p < 0.0001. Mo, morpholino; IOC, inner optic circle; NS, not significant.
|
