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Fig. 2. COUP-TFI-/- sensory epithelium exhibited extra
proliferating cells and ectopic hair cell differentiation in the supporting
cell region. (A-H) Immunofluorescence detection of p27 (red) to
mark the zone of non-proliferation (ZNP) in the sensory epithelia in wild-type
(A,C,E,G) and COUP-TFI-/- (B,D,F,H) cochlea at E15.5 (A-F)
and E17.5 (G,H). The ZNP was similar at the basal and middle cochlear turns
(A-D). However, the ZNP domain at the apex was widened (compare double arrows
in E with F) and encompassed extra hair cells (compare G with H) in the
COUP-TFI-/- mutants (myosin VIIa, green; GFI lectin I,
blue). The onset of hair cell differentiation was unchanged in the
COUP-TFI-/- basal (A,B) and apical (E,F) regions
(arrowheads point to myosin VIIa-positive cells). (I,J) BrdU
labeling (green) in wild-type (I) and COUP-TFI-/- (J)
epithelia at E16.5 shows extra positive cells in the outer sulcus region of
the apical turn in the COUP-TFI-/- mutants (arrows).
Sections were co-localized with myosin VIIa (red, arrowheads).
(K,L) p27 (green) expression in adjacent sections labeled with
BrdU, confirmed the BrdU-positive cells in outer sulcus region were outside
the ZNP. (M-P) Immunolocalization of Ki67 in wild-type (M,O) and
COUP-TFI-/- (N,P) cochlea at E16.5 (M,N) and P0 (O,P).
Proliferating cells were found in the outer sulcus region at E16.5 (red) and
inner sulcus region at P0 (green) in the apical turn of
COUP-TFI-/- mutants (arrows). (Q) Quantification of
BrdU-positive and Ki67-positive cells at E16.5. (R-T) Deconvolution
microscopy images show ectopic expression of myosin VIIa (asterisks in S and
T) in the supporting cell compartment of the COUP-TFI-/-
duct at P0. Scale bars: in A-P, 40 µm; in R-T, 100 µm.
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