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Fig. 3. Gli3 regulates proper establishment of the isthmus and
cerebellum. (A-E) RNA in situ hybridization for Math1 and
DAPI staining show that the external granule cell layer (EGL) is absent from
the most posterior and anterior parts (arrowheads) of the isthmus-cerebellar
like (Is/Cb) region in Gli3-/- mutants, but is comparable
to WT in En1-Gli3 cko mutants (arrowheads). (F-N)
Immunohistochemistry on adjacent sections. (F-K) In the WT, Purkinje cells
(PC) positive for calbindin and ROR
(green) are organized in a
several-cell-deep layer (PCL) underlying the EGL and project into the deeper
Cb (arrowheads). In Gli3-/- mutants, only a rudimentary
PCL forms with disorganized projections (arrowheads), and many PC remain in
clusters in the deeper Is/Cb (outlined). (H,K) In En1-Gli3 cko
mutants, most PC are located within the PCL, with only some scattered PC in
the underlying areas (H',K', arrowheads) and in ectopic clusters
in the anterior Is (outlined). Some PC axons project into the Is (H,
arrowhead). (L,M) Pax2 (green) is expressed in a scattered pattern throughout
the Is and Cb (except the EGL and PCL) in WT and En1-Gli3 cko
mutants, but is not expressed in the anterior (EGL-free) region in
Gli3-/- mutants and is excluded from the PC clusters (M,
arrowheads). (O-Q) H+E staining of P2 sagittal sections shows the
abnormal foliation pattern in En1-Gli3 cko mutants. Brain regions are
outlined where necessary. Note that some of the pictures presented are
composites of two images (C,D,F,G,I,J). Scale bars: 200 µm in A-N; 20 µm
in H',K'; 500 µm in P-Q.
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