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Fig. 7. NCCs populate epicardium and ventricular myocardium. (A-C)
Wnt1-Cre; R26R embryos showed blue
ß-galactosidase-positive cells (arrowheads) immediately caudal to the
heart (abutting the septum transversum) and on the cardiac ventral surface, in
the epicardium at E9.5 (A), E10.5 (B) and E11.5 (C). A transverse section of
the heart (C,F) reveals stained cells in the epicardium (epi) and myocardium
(m). ivs, interventricular septum; pp, parietal pericardium. (D,E) Neonatal
Wnt1-Cre; R26R hearts show ß-galactosidase-positive cells
(arrowheads) contributing to the coronary vasculature, consistent with an
epicardial lineage (inset shows entire heart shown in detail in D). (E)
Sections through E17 hearts (relative plane of section indicated in D inset)
indicate ß-galactosidase-positive cells surrounding an artery containing
red blood cells. (G,H) Similar ß-galactosidase-positive cells are seen in
the epicardium of PO-Cre; R26R embryos. (I) RT-PCR analysis to assay
for expression of Wnt1-Cre in ventral tissues. From each of two
Wnt1-Cre; R26R embryos (Cre+) and one control R26R embryo
(Cre–), RNA was isolated from tissues ventral to the dorsal aorta
(ventral), including the heart and septum transversum, and the neural tube
(neural). RNA was reverse-transcribed and subjected to PCR amplification of
Cre or Hprt (positive control). HPRT was amplified from all
samples except water (H2O) or those prepared without reverse
transcriptase (RT–). Cre was amplified only in the neural tube samples
from Cre+ embryos. (J-L) Bmp4-lacz expression. Whole-mount embryos at
E9.5 (J) and transverse sections at E9.5 (K) and E11.5 (L), showing high
levels of Bmp4 expression in the septum transversum (s.t.) and
parietal pericardium (pp), but not epicardium. cm, compact ventricular
myocardium.
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