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Figure 5


Fig. 5. Effects of BMP4 application at the optic vesicle stage (stage 8/9) on the distribution of genes expressed within the NR and RPE at stage 24. (A) Schematic illustrating the implantation site of the BMP4-soaked bead following the operations at stage 8/9. (B) Following BMP4 application into the optic vesicle, optic cup formation is not observed. Instead, a huge vesicle with RPE-like morphology, expressing Otx2 in both the proximal (C) and distal (D) regions developed. The arrowhead indicates Otx2 expression in the RPE of the contralateral, unoperated eye. Note that at this stage (stage 24), Otx2 transcripts are also observed in cells of the native NR (Bovolenta et al., 1997). This expression pattern is also seen in a small ventral region of the BMP-treated eye that has still NR-like morphology (arrow). (C) The entire optic stalk region has a single-layered RPE-like morphology expressing Otx2 (arrows) following BMP4 exposure. (D) Following BMP4 application, the region that normally develops into the NR has a RPE-like morphology and expresses Otx2 (arrows). (E) BMP4 application into the optic vesicle inhibits NR and optic stalk/nerve development and induces Mitf expression in the entire optic vesicle. Note that the entire eye has RPE-like morphology (arrow). In the unoperated, contralateral eye, Mitf expression is restricted to the RPE (arrowhead) and no transcripts are observed in the multilayered NR. (F) Mitf expression (arrows) in the optic stalk/nerve region that developed a single-layered morphology following BMP4 application. (G) Following BMP treatment, Mitf expression is observed in the region that normally develops into the multilayered NR (arrows). (H) Parallel section of the embryo shown in B. Chx10 expression is only detected in a small ventral portion of the BMP-treated eye (arrow). In the contralateral eye, strong Chx10 expression is restricted to cells of the NR (arrowhead). (I) The optic stalk/nerve region of the BMP-treated eye has RPE-like morphology and pigment granules (arrows) are observed. (J) Following BMP4 application, expression of the NR-specific marker Chx10 is not observed in the region that normally develops into the NR. Instead, this region is pigmented (arrows) and has RPE-like morphology. (K) Fgf8 expression is not observed in the distal region of the BMP-treated eye (arrows). (L) Following BMP4 application more temporal to the optic vesicle, eye morphology including lens development appears to be normal although the operated eye is slightly smaller. (M) MMP115 expression is restricted to the outer optic cup, the developing RPE and no transcripts are observed within the NR (arrows). (N,O) BMP4 application induced MMP115 expression in single cells of the NR (arrows). The asterisk indicates the neurepithelium of the diencephalon. L, Lens.





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