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Figure 2


Fig. 2. RMCE-mediated knock-in of lacZ into the Ednra locus. (A,B) Whole-mount staining of E8.25 (six-somite stage) (A) and E8.5 (nine-somite stage) (B) EdnralacZ2FRT/+ mouse embryos shows lacZ expression in the head mesenchyme at the hindbrain level (arrows). (C) An E8.5 transverse section at the rostral hindbrain level shows lacZ expression in mesenchyme throughout the head and pharyngeal arch regions, but not in the neuroepithelium, surface ectoderm and foregut endoderm. (D) High magnification of the boxed area in C demonstrates lacZ expression in migratory neural crest cells delaminating from the dorsal neuroepithelium (arrows). (E) At E9.0 (17-somite stage), lacZ is expressed in the head and pharyngeal arch regions, in the heart, and in the ventral half of the trunk. (F) An E9.0 sagittal section in the pharyngeal region demonstrates lacZ expression in neural crest-derived ectomesenchyme and in the head mesenchyme adjacent to the neuroepithelium, but not in the pharyngeal epithelium, vascular endothelium and many cells in the core mesenchyme (asterisks). (G) The E9.0 cardiac outflow region demonstrates lacZ expression in cardiac neural crest cells surrounding the second and third arch arteries and colonizing between the foregut and the aortic sac (arrow). (H-J) Whole-mount in situ hybridization of E9.0 wild-type embryos using Ednra (H), Crabp1 (I) and Snail1 (J) probes. (K-M) Comparison between Ednra-lacZ expression (K) and in situ hybridization patterns of Crabp1 (L) and Snail1 (M) in E9.0 transverse sections at the first pharyngeal arch level. White arrowheads, migratory neural crest cells; arrows, mesoderm-derived mesenchyme. Scale bars: 100 µm in C,F,G,K-M; 50 µm in D. aa2/3, second and third arch arteries; as, aortic sac; fb, forebrain; fg, foregut; hb, hindbrain; ht, heart; hv, head vein; mb, midbrain; ne, neuroepithelium; nt, neural tube; op, optic vesicle; ot, otic vesicle; pa1/2/3, first, second and third pharyngeal arches; pp2, second pharyngeal pouch.





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