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Fig. 1. Absence of mesencephalon in OtxH embryos. (A) Western blot
analysis on wild-type or OtxH zebrafish embryo extracts from end of
gastrulation stage (upper panel: bud stage) or late somitogenesis stage (lower
panel: prim-5). Proteins staining is shown underneath each to control protein
levels between wild-type and OtxH embryos. 1>, mouse Otx1protein; 2>,
doublet of mouse or zebrafish Otx2 proteins. The horizontal black line
represents the 36.4 kDa marker. In OtxH embryos, Otx2 expression is not
visibly affected at the end of epiboly but is lost by the end of
somitogenesis. (B-E) Lateral views, anterior towards the left, of
prim-20 (33 hpf) live brains (B,C) or fixed after acetylated tubulin staining
(D,E) from wild-type (B,D) or OtxH (C,E) embryos. The isthmic constriction
(arrow in C), the ventral flexure (VF) and the posterior commissure (PC,
normally forming at the boundary between forebrain and midbrain) are all
absent. tel, telencephalon; o.p., olfactory placode; tect., tectum; ist,
isthmus. Dorsal (F,G) and lateral (H-K) views, anterior
towards the left, of wild-type (F,H,J) or OtxH (G,I,K) brains. Expression of
mbx is almost undetectable in the CNS at the six-somite stage in OtxH
embryos (arrow in G) compared with wild type (arrow in F) and is lost by the
22-somite stage (I). Expression of Engrailed is detectable in both wild type
(J) and OtxH (K) at prim-5 stage. Insets are dorsal views of the isthmic area
of the brains shown in J and K.
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