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Fig. 3. Region-specific apoptosis and defective cell proliferation in the
RA-deficient forebrain. (A,B) Immunofluorescent TUNEL
detection of apoptotic cells on transverse forebrain sections of 14- to
15-somite stage wild-type (A) and Raldh2-/- (B) embryos.
Overlays of the DAPI nuclear staining (blue) and TUNEL signal (red) are shown
in false colors. (C,D) Whole-mount TUNEL analysis of wild type
(C) and mutant (D) E9.5 embryos. Frontal views of the head.
(E,F) Immunofluorescent detection of phospho-histone H3 on
transverse forebrain sections of 14- to 15-somite stage embryos (false color
overlays as in A,B). (G) Numbers of phospho-histone H3-positive cells
as percentages of total cell numbers along the whole ventral forebrain
neuroepithelium (fb) and three subregions, the rostroventral diencephalon
(di), optic vesicles (ov) and ventral telencephalon (te) of 14-somite stage
wild-type and Raldh2-/- embryos. Four embryos of each
genotype were sectioned transversely, and all sections encompassing the
aforementioned regions were counted (fb: wild type, 8%±1.3; mut,
5.1%±1.1; mean±s.e.m.; t-test P=0.0147; di:
wild type, 6.5%±1.7; mut, 3.3%±0.9; P=0.0158; ov: wild
type, 8.9%±1.6; mut, 5.4%±1.3; P=0.0131; te: wild type,
8.0%±2.0; mut, 6.3%±1.5; P=0.219). (H-M)
Whole-mount in situ hybridization of p21 (H), cyclin D2 (J,K) and
cyclin D3 (L,M) in wild-type and Raldh2-/- embryos. H,I:
ventral views of E9.5 heads; J-M: profile views of 13- to 14-somite stage
embryos. Insets show ventral (J,K) and frontal (L,M) views of the forebrain.
ma, maxillary process; me, mesencephalon.
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