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Fig. 7. FGF10 controls actin fiber formation (A-I), cell polarity (J-K) and
expression of vimentin (L-M) in the developing eyelid epithelium. Whole-mount
(A, D-G) and section (B,C,H,I) staining for F-actin of the eyes from
wild-type, Fgf10+/– and
Fgf10–/– fetuses at E15 (A-D,G-M) and E16
(E,F). The upper (B,H) and lower (C,I,J-M) eyelid primordia are shown. (A,D-F)
The accumulation of actin fibers at the eyelid leading edge is accelerated, as
eyelid closure proceeds. (B,C) Actin fibers are accumulated in the leading
edge cells (arrowheads). (G-I) The Rhodamine-phalloidin binding to F-actin is
much less on the eyelid margin and in the epithelial leading edge (arrowheads
in H and I) of Fgf10-null fetuses. (J-K) Immunofluorescence of
-tubulin. (J,K) The localization of -tubulin is indicated by the
red, dotted signals. The borders of the cells are shown in green
(Bodipy-ceramide). The expression of -tubulin in the inner basal layer
(along the dotted line; approximately 17 cells were assessed) appears
disorganized and reduced in the Fgf10–/–
eyelid (K). (J',K') Higher magnifications of the eyelid tips shown
in J and K, respectively. Weak expression of -tubulin is detected
apically in the mutant epidermal cells (arrowhead in K'). (L,M)
Immunofluorescence of vimentin. Note that high levels of vimentin protein are
expressed in the eyelid mesenchyme (m). (L) In the wild type, vimentin is also
localized in the leading edge epidermal cells. (M) Mutant eyelid epidermal
cells express much lower levels of vimentin. (L',M') Higher
magnifications of the eyelid tip shown in L and M, respectively. The
arrowheads indicate the expression of vimentin. All images except A,D-G, which
were obtained with a fluorescence stereomicroscope, were captured by laser
scanning confocal microscopy. Scale bars: 300 µm (A,D-G); 25 µm
(B,C,H,I); 25 µm (J,K); 50 µm (L,M).
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