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Fig. 8. BMP7 and SHH are sequentially required for Pax2 expression in the distoventral optic cup. Optic vesicle (A-E;A'-E') or optic cups (F-J) from wild-type (A-C,A'-C',F-H), Bmp7^-/- (D,E,D',E') or or^J (I,J) embryos were dissected and cultured as described in the Materials and methods in the absence (A,A',D,D',F,I) or presence (B,B',E,E') of BMP7, follistatin (C,C',G), SHH (J), or the 5E1 anti SHH blocking antibody (H). After 2 days in culture (2DIV) optic cups were fixed and hybridised with a Pax2-specific probe. Images show whole-mount views (A'-E',F-J) or cryostat sections (A-E) of same cultured cups. Addition of BMP7 does not significantly modify the expression of Pax2 in wild-type vesicles (B,B') but it rescues that of Bmp7-null optic vesicles (arrows in E). By contrast, follistatin completely abrogates Pax2 signal in the optic vesicle (arrow in C) but not optic cups (arrow in G) from wild-type embryos. SHH rescues Pax2 expression in or^J cups (arrow in J), while the 5E1 antibodies prevents expression (arrow in H). Asterisks in H-J indicate RPE remnants; in these vesicles, the lens is visible through the tissue. Scale bar: 50 µm in A-F; 100 µm in G,H; 200 µm in I,J.

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