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Fig. 3. Gene expression posterior to ZLI enables further subdivision.
Six3 (A), Arx (B) and Gad67 (C) expression (blue)
and Shh expression (brown) are compared at E12.5. Besides strong
expression of Six3, Arx and Gad67 in p3, there is
distinctive expression of Six3+ close to Shh (A,
arrow), Arx+ close to pial (B, arrow), and Gad67+
close to Shh (C, arrow). Expression of Gad67 is
observed only in the medial part of the Rim, which is marked by
Nkx2.2 shown by fluorescent in situ hybridization (D, arrow).
Six3 expression close to Shh overlaps with that of
Nkx2.2 completely (E, inset). No part of the Arx
expression domain overlaps with that of Nkx2.2 (F, inset).
Two-color in situ hybridization of Tal2 (blue) and Nkx2.2
(brown) reveals that Tal2 is expressed in the Rim VZ (G, inset).
Tal2 expression in the Rim VZ overlaps with that of Mash1
but not that of Shh, as shown by triple fluorescent in situ
hybridization. A closer view of ZLI is shown (H). Pitx2 (blue)
is expressed lateral to Shh (brown) (I, arrow). Triple
fluorescent in situ hybridization demonstrates no overlap of Six3
(green), Pitx2 (red) and Shh (blue) (J). Scale bar in
A: 500 µm for A-G,I; 250 µm for H,I,E-G, inset.
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