Figure 2

Cell tracing study of neural crest cells in chick embryos. a, Experimental schema for cell tracing of neural crest cells in the chicken embryo. Constructs were electroporated into the neural fold of stage 8 embryos at the midbrain and anterior hindbrain level before neural crest cells delaminate from the neural tube. b, Stereoscopic images of GFP fluorescence observed in the in ovo chick embryo at 6, 24, and 48 hours after introduction of GFP by electroporation into the neural fold. c-f, Confocal images of direct GFP fluorescence and immunostaining of HNK1 in the chick embryos at 6 hours (c, d) and 48 hours (e, f) after electroporation. The asterisks in c indicate the neural tube, while the arrowheads in c and e indicate the double positive cells magnified in d and f, respectively. g,h; Confocal images of direct GFP fluorescence in the olfactory mucosa of chick embryos at 13 days after electroporation. The boxes in g indicate the area magnified in h. Anti-fibronectin staining demarcates the mesenchyme of the olfactory mucosa and demonstrates the presence of a GFP⁺ neural crest cell in the olfactory epithelium. The asterisks in h indicate the nasal cavity. Nuclei were counterstained with Hoechst. Scale bars: (c, e) 100 μm, (d, f, h) 20 μm, and (g) 200 μm.