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Fig. 3. Increased proliferation in the preoptic region and hypothalamus of
cavefish embryos is Shh-dependent. (A-H) PCNA immunoreactivity in
transverse sections (surface fish, left; cavefish, right) at 60 hpf (A-D) and
5 days (E-H, high magnification) through the preoptic region (A,B,E,F) and the
hypothalamus (C,D,G,H). Note that proliferation in the dorsal diencephalon and
mesencephalon is identical in both populations. (I,J) Cavefish
embryos were treated with 20 µmol/l cyclopamine from 15 to 24 hpf, allowed
to develop until 48 hpf, and proliferation was assessed with
anti-phosphohistone H3 immunostaining (see Fig. S1 in the supplementary
material for pictures). The results are expressed as the density of
phospho-H3-positive cells (number of cells per surface area in arbitrary
units). See methods for details. * indicates a significant effect
(P<0.05, n=4 embryos; Student's t-test) when
compared with the ethanol-treated controls. Scale bars: 50 µm. ddi,
diencephalon; hyp, hypothalamus; mes, mesencephalon; po, preoptic region.
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