Fig. 3. Expression of X-Ptc-1, X-Ptc-2, X-Smo, Gli1, Gli2 and Gli3 on retinal sections. These panels show retina vibratome sections after whole-mount in situ hybridisation. The probes are indicated on the left of each series of panels. (A,D,G,J,N,S) Stages 28-30 embryos. (B,E,H,K,O,T) Stage 34-36 embryos. (C,F,I,L,P,U) Stage 40-41 embryos. (M,V) Dorsal CMZ. (Q,R) Ventral CMZ. (A) X-Ptc-1 expression is detected in the periphery of the optic vesicle (arrows). (B,C) X-Ptc-1 expression is still detected in the periphery of the retina albeit at a low level, including the RPE (arrows). (D,G) X-Ptc-2 and Gli1 are expressed in the presumptive RPE (arrows). (E,H) X-Ptc-2 and Gli1 expression is still clearly detected in the RPE (arrows). (F,I) A faint X-Ptc-2 and Gli1 expression is detected in some cells of the RPE (arrows). (J,N,S) Gli2, Gli3 and X-Smo expression is detected in the periphery of the optic vesicle (arrows). (K,O,T) Gli2, Gli3 and X-Smo expression is restricted to the periphery of the retina (arrows). (L,P,U) Gli2, Gli3 and X-Smo expression is restricted to the periphery of the CMZ where stem cells are present. Magnifications of these regions (M,Q,V) show that these genes are also expressed in the peripheral pigmented epithelium (arrowheads point to stained cells with a longitudinal shape that shows its belonging to the RPE). (R) When the RPE has been poorly bleached, we can detect Gli3-positive pigmented cells (arrowhead). Dorsal is towards the top. le, lens; NR, presumptive neural retina; RPE, presumptive RPE; CMZ, ciliary marginal zone. Scale bar: 60 µm for A-L,N-P,S-U; 20 µm for M,Q,R,V.

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