(Downloading may take up to 30 seconds.
If the slide opens in your browser, select File -> Save As to save it.)


Fig. 9. Effects of Notch1 ablation on glial cell formation. (A,B) Immunofluorescent analysis of GFAP expression in the vermis of adult mutant (A) and control (B) animals revealed no obvious differences in the glial cell population or density of Bergmann glia fibers, suggesting that although the size of the vermis is reduced, the proportion and distribution of glial cells is not substantially affected in the mutants. (C) Confocal analysis of midline sagittal sections of adult control (heterozygous Floxed Notch1, En2-Cre, R26R) cerebellum immunostained with anti-GFAP (green) and anti-ß-galactosidase (red) antibodies revealed co-expression in a population of Bergmann glial cells (arrowhead). The soma of a Purkinje cell expressing ß-galactosidase but negative for GFAP is also indicated (asterisk). (D) Quantification of GFAP ß-galactosidase double-positive cells on midline sagittal sections through the cerebellar vermis showed a highly significant (**P<0.01; Student’s t-test) reduction in recombined glial cells in the mutant (7±3%; homozygous Floxed Notch1, En2-Cre, R26R ({Delta}/{Delta}) compared with the control animals (66±6%; heterozygous Floxed Notch1, En2-Cre, R26R ({Delta}/wt)). (E) Isolation of glial cells from the vermis region of the cerebellum of P4 control animals and immunostaining with anti-GFAP (red) and anti-ß-galactosidase (green) antibodies supported the presence of glial cells derived from Cre-recombinase-expressing progenitor cells (arrowheads). ß-Galactosidase-expressing GFAP-negative, putative neurons are indicated (arrows in E). (F) Quantification of four individual experiments to identify GFAP ß-galactosidase-expressing cells in control ({Delta}/wt) and mutant ({Delta}/{Delta}) animals. The percentage of GFAP ß-galactosidase double-positive cells is shown, as is the total number of GFAP-positive cells in each culture (n) and the average of all four experiments (Avr.). Note that the total number of cells in the cultures from mutants was lower than that from control animals due to the 50% reduction in size of the mutant vermis at P4. GL, granule cell layer; ML, molecular layer; W, white matter. Scale bar in A, 0.5 mm for A,B.