Fig. 3. Lack of Purkinje and granule cell abnormalities in double null mutants. (A,B) Staining for calbindin does not reveal detectable differences in Purkinje cell dendritic trees at 3 months of age between control (A) and double knockout (B) samples. (C,D) Golgi staining shows that spines in double null mutant Purkinje cells are comparable with those of control Purkinje cells (arrows, C,D). Note that the Purkinje cells in A-D are from the antrior folia, which show the most prominent ectopic granule cell accumulation. (E-F) Calbindin staining of a double null mutant sagittal section (F) demonstrates the continuous Purkinje cell layer comparable with a control (E) at 2 weeks of age (arrows in E,F). Scale bars: 50 µm in A,B; 25 µm in C,D; 150 µm in E,F. (G) Double null mutant and control samples display comparable cell proliferation (n=3). Cultured vibratome sections were treated with BrdU and after 16 hours stained for the presence of BrdU-positive cells as described (Wechsler-Reya and Scott, 1999). (H) The IGL of folia III-V covers a comparable area both in controls and double null mutants (n=3). The area encompassing the IGL layers was analyzed using MetaMorphR software. (I) Double null mutant and control granule cell cultures contain similar comparable amounts of apoptotic cells (n=3). Cultures were maintained as described (Galli et al., 1995) and treated for 24 hours either with 25 mM KCl in the presence of serum (protected against apoptosis, blue columns) or with 5 mM KCl in the absence of serum (apoptosis induced, red columns). The number of apoptotic cells was analyzed using FACS as described (Spector et al., 1997).
