Pax2a is necessary for recruitment of hyaloid vasculature into the optic fissure. (A)In vivo 4D confocal imaging of Tg[kdrl:mCherry] pax2a+/+,+/− or pax2a−/− embryos. Time lapse series depicting the region of the OF (dotted white lines) and mCherry positive vasculature endothelial cells migrating through the fissure from 24-29 hpf. Scale bar = 10 μm. (B) Comparison of pax2a+/+,+/− and pax2a−/− vascularization during OF fusion, 24-48 hpf. 3D reconstructions of whole mount Tg[kdrl:mCherry] (red) pax2a+/+,+/− or pax2a−/− embryos with DAPI (blue) stained DNA. Scale bar = 50 μm. (C) Quantification of the number of mCherry positive cells from 3D confocal stacks within the region of the OF. Individual embryo results are depicted. ANOVA p < 0.0001. (D) Whole mount in situ hybridization comparing tln1 expression between pax2a+/+,+/− and pax2a−/− embryos at 28 and 32 hpf. tln1 signal within the OF (yellow arrowhead) appears reduced in Pax2a−/− embryos (red *). (E) qPCR analysis of tln1 expression from heads of pax2a+/+ and pax2a−/− embryos at 32 hpf confirms a reduction in tln1 expression.
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