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Fig. 6. Increased cell proliferation and Fgf8 upregulation after CNC
ablation in chick embryos. (A-C'') A combined in situ
hybridization for Myf5 (purple) and BrdU staining (red) in transverse
sections of stage 18 embryos in the trunk (A-A'') and BA2 region
(B-B'', control) or CNC-ablated embryo (C-C''). Arrowheads point to
Myf5 expression and BrdU staining in the muscle anlagen. (D) A
quantification analysis of proliferating myoblasts in branchial arches of
control and CNC-ablated embryos at 26 and 45 hours of incubation. Similar
results were obtained in three independent experiments and in each experiment
the bars represent counts from three adjacent sections.
*P<0.05, **P<0.01.
(E-G'''') A double immunostaining for Myf5 (red) and BrdU
(green). (E-E'''') A transverse section at the trunk level.
(F-F'''') A transverse section at the level of BA2 in control
embryos. (G-G'''') A transverse section at the level of BA2 in
CNC-ablated embryos. Merged BrdU/Myf5 images plus higher magnifications
(inset) are shown on the right. In situ hybridization for Fgf8 in
control (H,H') or CNC-ablated embryos
(I,I'). (J) RT-PCR results of CPM explants from
control or 100 ng/ml FGF8b-treated explants incubated for 3 days in culture,
n=2/2. (K) RT-PCR results of CPM explants from
control or CNC-ablated embryos. Whereas CPM explants underwent myogenesis in
control cultures, a reduction in myogenesis was observed following CNC
ablation (n=4/4). my, myotome; nt, neural tube; ov, otic vesicle; ph,
pharynx. Scale bars: in A and E, 0.15 mm for B,C,F,G and 75 µm for
A'-A'',B'-B'',C'-C'',E'-E'',F'-F'',G'-G'';
in H, 0.5 mm for H'-I'.
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