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Fig. 1. Comparison of Xiro1, Delta1, Serrate, Notch, Hairy2A and
Xslug expression. Embryos were fixed at late gastrula (stage 12.5-13)
or mid-neurula stage (stage 18-19), and double or single in situ hybridization
was performed for each gene. The stages and probes analyzed are indicated in
each figure. Anterior is towards the top, and the sections are shown with the
dorsal side towards the top. d, dorsal; v, ventral; orange arrowhead, neural
crest. (A-E) Comparison of Xiro1 and Xslug expression at the
late gastrula stage. (A) Initial visualization of a double in situ
hybridization of Xiro1 (green). Xiro1 has a dorsal or neural
domain of expression, and transcripts are also found in the preplacode domain
outside of the neural plate (star). (B) Section of an embryo stained as in A.
(C) Visualization of Xslug in purple in the same embryo as shown in
A, where Xiro1 expression is in green. As the dorsal and placodal
domain (star) are continuous, Xiro1 overlaps with the neural crest
territory (orange arrowhead). (D) Section of the embryo shown in C. (E) Higher
magnification of the box highlighted in C. The continuity between the neural
domain and the preplacode domain (star) is visible. (F-J) In situ
hybridization for Delta1 and Slug. (F,G) Late gastrula
embryos were sectioned and divided in two groups. One group was stained for
Delta1 (F) and the other for Xslug (G). Note that
Slug expression, in the neural crest (orange arrowhead), coincides
with the gap in the expression of Delta1 at the neural folds. (H,I)
Double in situ hybridization for Delta1 and Xslug; note that
the cells expressing Xslug (arrowhead) are surrounded by cells
expressing Delta1 (arrows). (I) Higher magnification of the box
highlighted in H. (J) Section of embryos along the plane indicated in I; note
that the cells expressing Xslug are surrounded by Delta1
expression in the deep layer of the ectoderm. (K) Double in situ hybridization
of Serrate (purple) and Xslug (green), note that
Xslug expressing cells (orange arrowhead) are surrounded by
Serrate expressing cells (arrows) in the anterior neural crest
region. (L,M) Expression of Notch can be seen in the neural plate and
it overlaps with Xslug expression at the border of the neural plate
(arrowheads). (N-P) Expression of Hairy2A (N,O: green in left panel
and purple in the right panel of N, and green in O) can be detected in the
entire neural fold, including the domain were Xslug is expressed
(orange arrowhead). The bracket indicates the region in which Slug
and Hairy2A expression overlap. The arrow (N,O) indicates the
expression of Hairy2A in the prospective posterior neural crest. (P)
The expression of both genes persists at the late neurula stage. A summary of
the expression of all these genes is represented as a whole mount in Q, and in
a section in R.
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