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Fig. 5. Diastema epithelium can inhibit transcription of Ptc1 and
Shh in the mandibular arch. (A-F) E12.5 mandibular explants following
24 hours of culture. (A,B,D,E) ß-Galactosidase staining marks
transplanted E11.5 tongue and diastema epithelium, over the left incisor tooth
germs. (C) Ptc1 expression is normal in the presence of tongue
epithelium. (F) Ptc1 is downregulated in the presence of diastema
epithelium. (G,H) Isolated E11.5 incisor explants following 24 hours of
culture. (G) In the presence of E11.5 tongue epithelium, Shh
expression is normal (right incisor, arrowhead), as compared with the control
(left incisor, arrow). (H) In the presence of E11.5 diastema epithelium,
Shh expression is lost (right incisor, arrowhead), as compared with
the control (left incisor, arrow). The Shh-blocking antibody 5E1 can
downregulate Shh transcription. (I) E11.5 mandibular explant cultured
for 24 hours in the presence of a 2H3 bead (left) and a 5E1 bead (right)
placed adjacent to sites of incisor development; in the right incisor adjacent
to the 5E1 bead, Shh is downregulated. (J-M) Isolated diastema
mesenchyme is able to transcribe Ptc1, but not Shh. E13.5
tongue explants following 24 hours of culture. (J,L) GFP-label marks the
transplant. (K) Transplanted diastema mesenchyme transcribes Ptc1.
(M) Transplanted diastema mesenchyme fails to transcribe Shh. (N)
Ectopic Ptc1 expression in isolated diastema mesenchyme following
separation of the incisor and molar regions prior to epithelial removal and
culture for 24 hours. Scale bars: in A, 600 µm for A,D; in B, 100 µm for
B,C,E,F; in G, 600 µm for G,H,J-M; in I, 600 µm for I,N.
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