Figure 6. Glia-independent neuronal defects in the Nf1^Math1CKO cerebellum are rescued by P0.5–P21 MEKi-treatment.

Western blot analysis was performed on P8 whole cerebellar lysates (A) or P6 cultured GCP lysates (A′) from control and Nf1^Math1CKO mice. (B) Cerebellar sections from P8 control and Nf1^Math1CKO mice were stained for NeuN and p-Erk. Arrows highlight the NeuN⁺p-Erk⁺ cells in the mutant EGL and ML. (C) Control and Nf1^Math1CKO mice were treated with vehicle or MEKi (5 mg/kg) from P0.5–P21 and analyzed at P21. Cerebellar sections were stained for Tbr2/NeuN, and imaged at low magnification (upper panels) and folium V (lower panels). (C′) The number of NeuN⁺ cells in the ML was quantified and compared. Of note, the yellow triangles represent 3 MEKi-treated mutant cerebella that still displayed cell clusters near the pial surface. Individual data points are presented, as well as mean ± SEM for each group. DAPI labels the nuclei. Scale bars: 50 μm.

DOI: http://dx.doi.org/10.7554/eLife.05151.015

Figure 6—figure supplement 1. Rapamycin treatment during neonatal stages does not rescue the neuronal defects in the Nf1^Math1CKO cerebellum, but causes adverse effects.

(A) Sections of P8 control and Nf1^Math1CKO cerebella were stained for p-S6 and NeuN. (B) Control and Nf1^Math1CKO mice were treated with vehicle or Rapamycin from P0.5–P21 and analyzed at P21. Arrows point to p-S6⁺NeuN⁺ cells, which are also shown in the inset. (C) Rapamycin-treated control and Nf1^Math1CKO mice exhibited significant cerebellar defects. Sections from vehicle- or Rapamycin-treated controls were stained for NeuN and p-S6. Arrows point to folia where there is abnormal neuronal clustering in the Rapamycin-treated cerebellum, but not in the vehicle-treated control. DAPI labels the nuclei. Scale bars: 50 μm.

DOI: http://dx.doi.org/10.7554/eLife.05151.016