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Fig. 6. The Alk3 CKO embryos exhibit defects in the cushion formation
in the AVC, but not in the OFT. (A) E10.5 wild-type (n=4)
and Alk3 CKO (n=4) embryos were sectioned sagittally and
stained with Hematoxylin and Eosin. In the control (wild-type) embryos,
trabeculae and endocardial cushions in the AVC region, as well as the OFT
develop normally. The AVC cushion is missing in the Alk3 CKO embryos,
but the development of the OFT cushion persists. Black arrows and white arrows
represent the cushion in the AVC and in the OFT, respectively. Middle and
bottom panels represent higher magnification of the AVC and the OFT,
respectively. Scale bars: 200 µm. (B) Flk1+/Cre
mice were crossed with Rosa26R-lacZ mice and the resulting embryonic
hearts were cross-sectioned, and subjected to lacZ staining. At
E10.5, the majority of lacZ-positive cells in the AVC are present in
the cushion cell layer and in the endocardium. However, in the OFT, the
lacZ-positive cells are limited to the endocardium. By E12.5, all ACV
cushion cells are lacZ+, while the OFT cushion consists of
both lacZ+ and lacZ- cells. (C)
Alcian Blue staining. (D) Quantitative RT-PCR analysis of E10.5 heart
lacking the OFT and right ventricle for Snai1, VE-Cadherin (VE-Cad),
Sox9, Nfatc1, Tgfb2, Msx1 and Twist1. Four wild-type and four mutant
embryos were used for quantitative RT-PCR analysis.
*P<0.05.
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