Zebrafish med14 mutant recapitulates the phenotypes of VACTERL association. (IMAGE)
Caption
(A) med14 mutants showed heart looping defects. At 48 hpf, heart tube looping was assessed in med14 mutants or WT with a Tg(cmlc2-kalta4-p2a-mCherry) background (left) or using whole-mount in situ hybridization (WISH) with a cmlc2 probe (right). The ratios of affected embryos were indicated. Scale bar, 50 μm. (B) Alcian blue staining in the WT and med14 mutants from 3 dpf to 5 dpf. Scale bar, 100 μm. (C) Alcian Blue and Alizarin Red S staining of WT and med14 mutants at 9 dpf. The ratio at the bottom-right corner represents the number of embryos with the indicated phenotype versus the total number of embryos examined. (D) Live images of cloacae in the WT and med14 mutant embryos in the lateral view at the indicated time points. The ratio at the bottom-right corner represents the number of embryos with the indicated phenotype versus the total number of embryos examined. (E) Immunostaining for α-PKC in WT and med14 mutants. Nuclei were counter-stained with DAPI. Scale bar, 30 μm. (F) Rhodamine dextran excretion assay in med14 mutants. Rhodamine dextran was injected into the anterior gut of med14 mutants or WT. The movement and excretion of fluorescent dextran from the gut were monitored 8 h post injection (also see Videos S1, S2). (G) Assessment of kidney function in med14 mutants. WT and med14 mutants were injected with 40-kDa rhodamine-labeled dextran via the cardiac venous sinus. The fluorescence intensity of each fish was measured at baseline and after 1, 5, and 24 h.
Credit
Jingwen Liu, Xuelai Liu, Feifei Li, Yujie Wang, Liping Yang, Meijia Zhang, Xinyu He, Ting Zhang, Ying Yi Zhang, Qiang Wang
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