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Fig. 3. Assessment of Pax8 function in otic vesicle patterning. (A,E) Otic vesicles
at 32 hpf stained with anti-Pax2 (red) and anti-acetylated tubulin (green)
antibodies to label hair cells (Riley et
al., 1999) in wild-type (A) and pax8-MO-injected (E)
embryos. Hair cell patches are indicated (white arrows). Injected embryos
produce fewer hair cells than normal. In the experiment shown here, an average
of 5.2±1.1 hair cells were seen in pax8 morphants
(n=59 ears), compared to 9.2±1.4 hair cells in wild type
embryos (n=10 ears); 5-10% of pax8 morphants produce only
one macula per ear (not shown). (B,F) pax5 expression at 24 hpf in
wild-type (B) and pax8-MO-injected (F) embryos. (C,G) dlx3b
expression at 33 hpf in wild-type (C) and pax8-MO-injected (G)
embryos. (D,H) msxC expression marks developing cristae in the otic
vesicle at 78 hpf in wild-type (D) and pax8-MO-injected (H) embryos.
The lateral crista is present in the injected embryo but shows strongly
reduced expression of msxC. Defects in development of cristae and
semicircular canals are highly variable in pax8 morphants. Images
show lateral views with anterior to the left and dorsal to the top. ac,
anterior crista; lc, lateral crista; pc, posterior crista. Scale bar in H: 90
µm for D,H, 40 µm for C,G, and 30 µm for A,B,E,F.
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