Fig. 4. Induction of the CMZ markers and inhibition of neural retinal markers by Wnt2b overexpression in ovo. (A-H) The expression pattern of NF-M (A,B), Notch1 (C,D), Pax6 (E,F), Chx10 (G,H), Rx1 (I,J), and collagen type IX (K,L) in the embryos electroporated with control (A,C,E,G,I,K) or chicken Wnt2b (B,D,F,H,J,L)-expressing plasmids. The embryos were fixed at stage 21, and adjacent transverse sections were stained for co-electroporated GFP (shown in left panels) and each molecular marker (shown in right panels). In the Wnt2b-electroporated embryos, the inner layer of the optic cup formed folded retinas that did not express neural retina-specific markers such as NF-M or Notch1 (B,D). The folded retinas expressed progenitor marker genes, including Pax6 (F), Chx10 (H) and Rx1 (J). Note that whole retina was affected by relatively uneven introduction of the electroporated chicken Wnt2b. Arrowheads indicate the central neural retina. di, diencephalon. Scale bar: 100 µm.