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Fig. 9. Msx1/2 are required for the survival of cranial neural
crest cells. (A-F) Double-label immunostaining of apoptotic and proliferating
cells. Immunostaining of phosphorylated histone H3 (Rhodamine, red) and TUNEL
(FITC, green) were carried out on the same sections to detect cell
proliferation and apoptosis in Msx1/2 mutants. Nuclei were
counterstained with DAPI. Arrowheads indicate increased apoptotic neural
crest-derived cells in the region of the trigeminal ganglion (B), the proximal
portion of pharyngeal arch1 (maxillary prominence; D), and the distal tip of
pharyngeal arch1 (mandiblular prominence; F) of Msx1/2
mutants at E9.5. Cell proliferation within these sites was not noticeably
altered. (G-L) Nile Blue-stained E9.5 control (G,I,K) and
Msx1/2 mutant (H,J,L) embryos; lateral (G-J) and dorsal
(K,L) views. Increased cell death was detected in neural crest-derived
craniofacial structures of the Msx1/2 mutant (arrows,
arrowheads and open arrowheads in H and J) consistent with results of the
TUNEL assay. We did not detect increased cell death in cardiac neural crest
cells in mutant embryos (J). Note the lack of discernable change of cell death
in the mutant hindbrain (L). Scale bar in L: 0.2 mm (for G-L).
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