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Fig. 3. Shh::GFP ligand forms a gradient at the apical region that can be
modified by Smo and Skn activity. (A-I) Sections
of 14- to 15-somite-stage mouse neural tubes of indicated genotypes. (A-C)
Immunostaining for Nkx6.1, whose expression is activated by Shh signaling, and
for Pax6, whose expression is repressed by Shh signaling in the ventral neural
tube. (D-F) Shh::GFP ligand distribution directly visualized by confocal
microscopy. (G-I) High magnification of the apical, ventral neural tube.
Images are a projection of six confocal images taken sequentially along the
z-axis compressed into one to provide a better sampling of the total
Shh::GFP ligand distribution. Note that Shh::GFP ligand forms a large punctum
near the ventral midline (G, red arrow) at this stage that is presumably made
up of several smaller puncta. White arrows indicate approximate dorsal limit
of detectable Shh::GFP ligand. (J) Profiles of apical Shh::GFP ligand
along the D-V axis of indicated genotypes. Profile represents an average based
on two sections per embryo across three embryos. Approximate positions of
Shh-dependent progenitors p3, pMN and p2 in a ShhGFP/GFP
neural tube at this stage are displayed along the x-axis. Scale bars:
5 µm in G-I.
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