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Fig. 3. Shh is expressed in EZ-I and can rescue the mesethmoid loss induced by EZ-I ablation. (A-C) This HH14 chick embryo simultaneously received two quail grafts at the 5-somite stage. One was a homotopic substitution of the anterior neural crest and the other a supernumerary EZ-1 graft in the mesenchyme of the presumptive first PA. (A,B) Frontal section analysed by in situ hybridisation with a Shh probe, showing strong expression in the floor plate, in the notochord and in the grafted EZ-I (boxed). (C) Immunodetection of quail nuclei confirms the quail origin of the grafted endoderm and of the neighbouring CNCCs. (D) Dorsal schematic view of a 5-somite chick embryo showing the strategy of Shh rescue. After EZ-I ablation (Step 1), a Shh-loaded bead was implanted in the vacant region (Step 2). (E) Representative rescued embryo (HH35) showing almost normal development of the mesethmoid. (E') The same embryo before skeletal preparation showing the development of an upper beak. 1^stpa, first pharyngeal arch; I and II, endoderm zones I and II; ect, ectethmoid; en^*, supernumerary graft of EZ-I; fp, floor plate; Mc, Meckel's cartilage; mes, mesethmoid; n, neural tube; nt, notochord; QNCC, quail neural crest cells; r1-r8, rhombomeres 1-8.

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