|
Fig. 4S1
Additional diencephalon phenotypes in fezf2 mutant embryos.
(A–F) Overlay analysis of in situ expression patterns in wild-type and fezf2 mutant embryos at 2 and 3 dpf. Wild-type expression patterns are shown in green and fezf2 mutants are shown in magenta. Expression of the GABAergic markers dlx5a (A) and gad1b (B) begins to recover in the ventral diencephalon (arrows) of mutant embryos at 3 dpf (compare with Figure 4A,B). (C) th expression is significantly reduced in the dopaminergic (DA) clusters of the ventral diencephalon, with posterior clusters (open arrowheads) appearing to be more strongly affected than anterior clusters (arrows). (D) Expression of the otpb transcription factor is lost in the ventral posterior tuberculum and posterior hypothalamus (arrows) but persists in other brain regions. (E) Vmat2 expression is lost in DA clusters of the ventral diencephalon (arrows). (F) The fezf2 expression domain is dramatically altered in the posterior hypothalamus (arrow) and is substantially reduced in the retina at 3 dpf (but note that retinal expression appears normal at 2 dpf; Figure 4C) (G–H) Multi-probe co-expression analysis of ventral diencephalic deficits in fezf2 mutants. (G) Virtual co-registration showing spatial relationships between affected otpb, th, and vmat2 expression domains in the ventral diencephalon at 3 dpf. (H) The sst1.1-expressing clusters lost in fezf2 mutants (arrow) partially overlap with optb in the hypothalamus and are either intermingled with or located just lateral to th-expressing DA clusters. (A–H) All 3D reconstructions are generated by averaging eight or more embryos per experimental group. The position and orientation of each 2D slice within the embryo is indicated on the Nissl-stained two photon reference image to the right of the panel. Scale bar: 100 μm.