Figure 1.

NF1^hGFAP mice exhibit abnormal cerebellum. (A) X-Gal staining of a 1-mo-old hGFAP-Cre Rosa26^LacZ mouse cerebellum. Inset is a high-magnification view of folium IX. Note that X-Gal staining is present in the GCL but not the PL. Bars: 1 mm; inset, 100 μm. (B) Immunofluorescence staining for cre-recombinase expression, as indicated by GFP, in the neuronal and glial lineages of a 1-mo-old hGFAP-cre;Rosa26^GFP reporter mouse cerebellum. Bar, 100 μm. (C) Western blotting and quantification of NF1 in control and NF1^hGFAP whole cerebellar lysates. Molecular weight markers (in kilodaltons) are shown. Mean ± SEM; n = 3; (*) P < 0.05. (D) Representative whole-mount images of control and NF1^hGFAP brains. The cerebellum has been circled by red dashed lines. (E) Analysis of the brain weight to body weight ratio of control and mutant mice. Mean ± SEM; n = 5; (**) P < 0.01. (F) H&E staining of cerebellar sections from control and NF1^hGFAP mice shows dysmorphology in mutant mice. Red arrows indicate folium VII at P28. Higher magnification of folium VII is shown in the adjacent right panels. Note the presence of abnormally retained cell clusters at the pial surface (EGL) of the mutant cerebellum (black dotted line boxes are magnified in the panels at the far right). (G) Immunohistochemistry of control and NF1^hGFAP mouse sections with NeuN (granule neuron), GABAARα6 (granule neuron), calbindin (Purkinje neuron), and GFAP (BG) antibodies indicates defective folia layering and gliosis. Bar, 100 μm.