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Fig. 7. Expansion of the Shh-positive floor plate in wild-type and Lp/+
embryos following ROCK inhibition in culture. (A-H) Wild-type embryos;
(I-L) Lp/+ embryos. (A) Untreated (control) wild-type embryo
after successful Closure 1 (arrow). (B) Wild-type embryo treated with
10 µM Y27632 (Inhib), showing failed Closure 1 (arrows: entirely open
neural tube). (C,D) H&E staining of transverse sections at
the level of the dashed lines in A and B, respectively. Note the normal,
V-shaped neural fold profile in the untreated embryo (C), by contrast to the
broadened floor plate region of the open neural tube after Y27632 treatment
(white arrow in D). (E,F) Whole-mount in situ hybridisation for
Shh in wild-type embryos cultured in absence (E) or presence (F) of
10 µmol/l Y27632. Shh expression domain is broader at the site of Closure 1
(dotted lines) in the treated embryo (arrow in F) than in untreated control.
(G,H) Transverse sections of Shh whole-mounts, at the
level of the dotted lines in E and F, respectively. Shh expression
domain is markedly expanded in the inhibitor-treated embryo (arrow in H).
(I-L) Sections of Lp/+ embryos cultured in absence (C) or
presence (D) of 2.5 µmol/l Y27632, followed by in situ hybridisation for
Shh. Expression is detected mainly in the notochord (n) of the
untreated control embryo (I; higher magnification in K), with a few positive
floor plate cells (white arrow in K). In the inhibitor-treated embryo (J;
higher magnification in L), Shh is expressed similarly in the
notochord but also in a laterally expanded floor plate domain (fp). Scale
bars: in F, 0.4 mm for A,B,E,F; in C, 0.1 mm for C,D; in G, 0.05 mm for G,H;
in I, 0.1 mm for I,J; in K, 0.025 mm for K,L.
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