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Fig. 6. Fgf8 is required for cell survival in the neurogenic domain. (A)
In situ hybridization for Fgf8 ex1 probe indicates areas where
(nonfunctional) Fgf8 is expressed in mutants (Fgf8 ex2,3
probe signal was not detected in mutants). TUNEL panel shows high number of
apoptotic cells in mutants in ectoderm (white asterisk) and OE (white
arrowhead; magnified in inset) of invaginating nasal pit (NP). Hoechst panel
shows extent of invaginating NP. LNP, lateral nasal process; MNP, medial nasal
process. Sox2 expression is reduced in OE of MNP (black arrowhead).
Scale bars: 100 µm. (B) High-power micrographs of TUNEL staining in OE of
E12.5 Type B mutant and control littermate. Broken white line indicates basal
lamina (BL) of OE. AL, apical surface; Str, stroma. Scale bar: 50 µm. (C)
Total TUNEL+ cells in identifiable nasal epithelium (NE) were counted, and
area of NE measured, in multiple sections at each age indicated. Data for each
NP were summed and normalized to 0.1 mm2, the average total NE area
in a section at E12.5. Mean±s.d. for data from individual NPs are
shown; NPs from a minimum of 2 animals of each genotype at each age were
counted. Differences between mutants and controls were statistically
significant at E10.5 (P=0.009, Student's t-test) and E12.5
(P=0.003), but not E14.5. (D) High-power micrographs of
anti-phosopho-Histone H3 immunostaining at E10.5. Scale bar: 50 µm. (E)
Quantification of data illustrated in D. (F) High-power micrographs of
anti-BrdU immunostaining at E10.5. Scale bar: 50 µm. (G) Quantification of
data illustrated in F. There are no significant differences between datasets
in E and G. Data for each NP were summed and normalized to 0.03
mm2, the average total OE area in a section at E10.5.
Mean±s.d. for data from individual NPs is shown; NPs from a minimum of
two animals of each genotype at each age were counted.
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