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Fig. 5. Analysis of Gbx2, Otx2, Hoxa2 and
Pax6 expression and neuronal death in the MHB of
Lmx1b-/- embryos. (A-F) Gbx2
expression was normal in mutant embryos at E7.75 (B), but was barely
detectable in the mutant brain at the 4-somite stage (D, arrow) and lost
altogether in the isthmus at E9.5 (F, arrow). Note that Gbx2
expression was normal in the caudal hindbrain (D, arrowhead) and otic vesicle
(F, arrowhead), as compared with controls (C,E). (G-J) Expression of
Otx2 was not changed in mutant embryos (H,J) as compared with
wild-type controls (G,I). (K-N) Double in situ hybridization for
Otx2 and Hoxa2 showing no changes in the distance
(double-arrowed line) between Otx2+ and
Hoxa2+ domains at the 5-somite stage (L) and a reduction
in this distance in the mutant embryos at the 13-somite stage (N), as compared
with controls (K,M). (O,P) Expression of Pax6 was not
changed in the mutant embryos (P) as compared with wild type (O). Note that
the rostral-caudal distance (double-arrowed line) marked by Pax6 was
maintained in the Lmx1b-/- embryo. (Q-T)
There was a slight increase in the number of TUNEL+ cells (arrow)
in the MHB in the mutant embryo at the 7-somite stage (R), when marked
downregulation of gene expression was observed. However, there was a
significant increase in the number of TUNEL+ cells (arrow) in the
MHB of the mutant embryo at the 13-somite stage (T), as compared with the
wild-type embryo (S).
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