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Fig. 3. Morphological comparison of Ptf1a^+/Cre⁽^ex1⁾ R26R and Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinae. (A,E) To confirm the absence of Ptf1a protein in Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinae, retinal sections from E18.5 embryos were immunostained with an antibody against Ptf1a (arrows). (E) No immunoreactivity can be detected in Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retina. (B,C,F,G) X-Gal staining of Ptf1a^+/Cre⁽^ex1⁾ R26R and Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinae at E18.5. Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retina shows morphological change in the inner retina (F) compared with the Ptf1a^+/Cre⁽^ex1⁾ R26R retina (B). Higher magnification of boxed areas in B and F are shown in C and G, respectively. At E18.5, the GCL and IPL are formed in Ptf1a^+/Cre⁽^ex1⁾ R26R retina (B,C). ß- galactosidase-expressing cells are primarily restricted to the inner zone of the NBL (C, arrows). By contrast, in Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retina (F,G), the GCL and NBL are fused and the IPL is lost. The ß-galactosidase-positive cells are scattered in the inner retina (G, arrows). In both Ptf1a^+/Cre⁽^ex1⁾ R26R and Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R retinae some isolated cells are found in the outermost zone of the NBL. (B,C,F,G) Background is stained by neutral red. (D,H) Images are taken in ApoTome mode (Zeiss) for the purpose of 3D reconstruction showing immunohistochemistry for N-cadherin (red) with a DAPI counterstain (blue). (D) In Ptf1a^+/Cre⁽^ex1⁾ R26R retina, N-cadherin stains GCL and IPL. (H) By contrast, N-cadherin-positive processes are strongly reduced in Ptf1a^Cre⁽^ex1⁾^/Cre⁽^ex1⁾ R26R, resulting in loss of the IPL. Scale bars: 50 µm in A,C,E,G; 100 µm in B,F; 10 µm in D,H. GCL, ganglion cell layer; IPL, inner plexiform layer; NBL, neuroblastic layer; rpe, retinal pigment epithelium.

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