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Fig. 3. In situ hybridization analyses of Ext2–/–
mutants. Hybridization was performed with 35S-labeled riboprobes
and sections were counterstained with Hematoxylin (A-J) or Hoechst (K-T). In
all sections, embryos are viewed laterally and the anterior side of the embryo
is towards the left. (A-H) Embryos collected at E6.5. (A,B) H19, a marker for
extra-embryonic tissues shows the presence of extra-embryonic ectoderm,
endoderm, ectoplacental cone and trophoblast giant cells in
Ext2–/–. The broken line indicates the
difference in development of the extra-embryonic parts of the egg cylinder.
(C,D) Apoe, a marker for visceral endoderm, is expressed in
Ext2–/– embryos. (E,F) Fgf8, a marker for
primitive streak formation, is detected in wild-type embryos and in
Ext2–/–. (G,H) Brachyury (T), another marker
for primitive streak and mesodermal cells, is present in wild-type embryos,
but not in Ext2 mutants. Embryos collected at E7.5 (I-T). (J) Class 2 mutant;
(L,N,P,R,S,T) class 3 mutants; (I,K,M,O,Q) wild type. Brachyury is expressed
in class 2 mutants (J) and in class 3 mutants (L,N,P,R,S,T). (M,N) H19 was
used as a probe to show the boundaries between embryonic and extra-embryonic
regions of the wild-type embryos and Ext2–/–
mutants. (O,P) Lim1 is expressed in wild-type embryos at low levels
in primitive streak and at higher levels in mesodermal cells migrating away
from the streak. Arrowheads indicate several cells expressing Lim1 in the
Ext2–/– mutants. (Q,R) Snai1
expression is shown in the primitive streak and nascent mesoderm of wild-type
embryos (Q) but is absent in Ext2–/– embryos
(R). (S) Otx2 and (T) Hesx1 mRNAs are detected in the
anterior cell mass of class 3 Ext2 mutants. ecp, ectoplacental cone;
eec, embryonic ectoderm; ps, primitive streak; tgc, trophoblast giant cells;
ven, visceral endoderm. Scale bars: 50 µm in A-H; 100 µm in I-T.
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