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Fig. 7. Overexpression of Fgf10 leads to the formation of apical bud in molar
germs. Stereomicroscopic observation (A-C) and frozen sections (D-F) of
cultured molar germs transfected with Fgf10 cDNA. (A) The
molar germ was cultured for 2 days after the transfection of Fgf10
cDNA expression vector. Fgf10 expression was identified by expression of green
fluorescence protein using a fluorescence stereomicroscope and was restricted
to the proximal dental papilla cells (arrow). (B) The molar germ was
cultured for 5 days. The expression expands along with the growth of the
dental epithelium. (C) Stereomicroscopic observation of the cultured
molar germ. The proximal dental epithelium (arrow) elongates and extends to a
greater extent than the distal dental epithelium (control side). (D)
CK14 immunostaining (red) of a frozen section clearly shows the formation of
an apical bud (larger arrow) and an expanded inter cuspal epithelium
(arrowheads) following Fgf10 overexpression. However, at the control
(distal) side, the epithelium forming the HERS consists of two or three
epithelial cell layers (smaller arrow). (E) The expression of Fgf10
(green) is seen in the proximal dental papilla cells between the proximal
dental epithelium and inter-cuspal epithelium. (F) The section was
counterstained with DAPI (blue) and the three colors were merged. (G)
Schematic illustration of the effects of Fgf10 overexpression. After
the morphogenesis of crown, Fgf10 mRNA disappears and the formation
of HERS starts (Fig. 1A).
However, at the proximal side, ectopic overexpression of Fgf10 protein leads
to apical bud formation. Conversely, at the distal side (control), HERS forms
as Fgf10 disappeared. Scale bars: 20 µm.
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