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Fig. 4. Disorganized vessel structure in the Alk3 CKO dorsal aorta.
(A-D') Anti-PECAM (CD31) antibody staining. The Alk3 CKO
embryos often exhibited dilated vessels (indicated by arrowheads) in the brain
and abnormal patterns of vessel branching in the trunk (B,D,D'), compared with
the controls (A,C,C'). (C',D') Higher magnification of the
insets of C and D, respectively. Scale bars: 1 mm. (E-J) E10.5 embryos
were stained with anti-PECAM1 (CD31) antibody (red, F,I) to visualize
endothelial cells and anti-SMC 
-actin antibody (green, G,J) to
visualize SMCs. The endothelial cell layer of the dorsal aorta appeared to be
normal, as indicated by anti-CD31 staining in both controls (wild-type) and
Alk3 CKO embryos. SMCs are readily found throughout the dorsal aorta
in the controls, but in the mutants SMCs are lacking in some areas of the
dorsal aorta. Arrows (E,H) indicate blood cells in the lumen of the dorsal
aorta. Three wild-type and three Alk3 CKO embryos were analyzed in
cross-section. Scale bars: 50 µm. (K) Quantitative RT-PCR analysis
of dorsal aorta for Cd31, SMC -actin, Mhc, Angpt1, Tie2,
Vegf, Pten, integrin v and integrin ß3. Three wild-type and
three Alk3 CKO embryos were used for qRT-PCR.
*P<0.01. (L) E10.5 embryos were sectioned and
examined under transmission electron microscopy. There are breaks in the
Alk3 CKO dorsal aorta (red arrowheads). In addition, the mutant
endothelial cell layer did not form a close association with SMCs, compared
with the controls. Two control embryos and four Alk3 CKO embryos were
analyzed for EM study. SMC, smooth muscle cells; Endo, endothelial cells.
Scale bars: 6 µm.
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