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Fig. 6. Effect of Shh on regeneration. Inhibition of the Shh pathway using KAAD increases the domain of transdifferentiation (A-C,J-L,S-U) compared to eyes that received Fgf2 alone (D-F,M-O,V-X). However, overexpression of Shh using Rcas-Shh decreases transdifferentiation (G-I,P-R,Y,Z). (A-I) E7. (A) At E7, KAAD/Fgf2-treated eyes show regeneration from the CB/CMZ (cr) as well the transdifferentiation of RPE (td). (B) A close up view of CB/CMZ from an E7 KAAD/Fgf2-treated eye. (C) A close up view of transdifferentiation from an E7 KAAD/Fgf2-treated eye. (D) Regenerating eye that has been treated with Fgf2 only. (E) A close up view of CB/CMZ regenerated retina. (F) Close up view of transdifferentiated retina. (G) Regenerating eye that has been treated with Rcas-Shh/Fgf2 shows almost no transdifferentiation. (H) CB/CMZ regenerated retina, where Shh has been overexpressed does not look different from that of E7 Fgf2-treated regenerating neuroepithelium (E). (I) Occasionally, there are small areas of transdifferentiation in the regenerating eyes that received Rcas/Shh/Fgf2. The neuroepithelium at this stage is similar to that of an Fgf2-treated transdifferentiating retina (F). (J-R) E11. (J) At E11, KAAD/Fgf2-treated eyes show an increased domain of transdifferentiation when compared to Fgf2-treated eyes of the same stage (M). (P) At this stage, several eyes that overexpress Shh show some transdifferentiation. KAAD/Fgf2 and Fgf2-treated eyes show similar organization having a GCL, IPL and INL (K,L,N,O); whereas Rcas-Shh/Fgf2-treated eyes seem to be more differentiated with a visible OPL and ONL (Q,R). (S-Z) E15. (S) At E15, most of the retina in KAAD/Fgf2-treated eyes results from transdifferentiation. However, there exist small regions that contain CB/CMZ regenerated retina. (V) Eyes treated with Fgf2 show both transdifferentiated retina and CB/CMZ regenerated retina. (Y) Rcas-Shh/Fgf2-treated eyes do not show any transdifferentiated retina and all the retina present is produced by regeneration from the CB/CMZ. All the retinas at this stage show similar levels of spatial organization (T,U,W,X,Z). Scale bars: 500 µm (A,D,G,J,M,S); 100 µm and is applicable to B,C,E,F,H,I,K,L,N,O,Q,R,T,U,W,X,Z; in P is 500 µm and applies to Y; 500 µm (V). An asterisk marks an Fgf2-coated heparin bead.