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Fig. 1. GFP labelling of cardiac progenitors in HH3 primitive streak.
(A) Schematic illustration of the electroporation and grafting
procedure; embryos were placed in EC culture with their ventral side up.
(B) HH3 donor embryo (left) after electroporation of primitive streak
with pCS2+-GFP, and HH3 host embryo (right). Green fluorescence
identifies labelled cells; small grafts were dissected and transplanted into a
stage-matched host. The white dotted semi-circles show the anterior margin of
the primitive streak; the white dotted oval indicates the graft. (C)
Transverse cryosection (15 µm) through an HH5 embryo shows GFP-positive
cells in the mesoderm of the lateral plate. (D) HH7 embryo with
GFP-positive cells in the area of the bilateral heart fields, indicated by a
white line (Stalsberg and DeHaan,
1969). (E) GFP-positive cells in the heart tube at HH10.
(F) Transverse cryosection (30 µm) of a similar embryo, stained with
anti-GFP (green) and MF20 (red) antibodies, showing that GFP-positive cells
were present in endocardium and myocardium. White line in E indicates the
approximate level of the section shown in F. ao, area opaca; ec, ectoderm; en,
endoderm; end, endocardium; fg, foregut; h, heart; hf, heart field; my,
myocardium; nt, neural tube; ps, primitive streak.
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