The effect of rnf2 deficiency on the migration of enteric neural precursor cells in zebrafish embryos. (A) The expression of phox2b in rnf2–/– and WT embryos at 24 hpf. Red arrows showing the reduced expression of phox2b in intestinal bulb of mutant embryos. Scale bar = 0.1 mm. (B) The expression of phox2b in rnf2–/– and WT embryos at 48 hpf. Black arrows showing the reduced expression of phox2b in the gut of mutant embryos, red arrows showing the ectopic expression of phox2b in the brain regions. The dashed black box showing the compromised migration of ENPCs. Upper: lateral view; bottom: dorsal view. (C) The expression of ret and hand2 in rnf2–/– and WT embryos at 48 hpf. Black arrows showing the reduced expression of hand2 in the gut of mutant embryos; red arrows showing the reduced expression of ret. The dashed black box showing the compromised migration of ENPCs. Upper: lateral view; bottom: lateral view. (D) The expression of Phox2b-GFP in the brain regions of rnf2–/– and WT embryos at 24 hpf (40 × oil, lateral view). The numbers of samples were 28 and 24 in 24 hpf WT and rnf2–/– embryos, respectively. The red dashed box showing the expansion of GFP signals. (E) The expression of Phox2b-GFP in gut regions of rnf2–/– and WT embryos at 96 hpf (40 × oil, lateral view and head is to the left). The white dashed box showing the different distribution of GFP+ signals in the gut of WT and rnf2–/– embryos at 96 hpf. The numbers of samples were 30 and 27 in 96 hpf WT and rnf2–/– embryos, respectively. (F) Quantitation of GFP positive cells in (E). The star indicates significant differences at p ≤ 0.05. The experiments were repeated at least three times. Scale bar: 0.1 mm.
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