Fig. 4: Tumor progression recapitulated in a clinically relevant glioma mouse model.
a Schematic illustration of experimental design for glioma induction and measurements in the glioma mouse model. Deletion of PTEN, Ink4a/Arf, and overexpression of K-RasV12 were induced by AAV-GFAP-cre injection in the primary motor cortex of triple conditional mouse. Measurements were carried out on 7, 14, and 21 days post injection (DPI). Brain outlines from Allen Mouse Brain Atlas (atlas.brain-map.org). b Representative hematoxylin-eosin (HE) staining and Ki67 immunolabelling of mouse brain with tumor-bearing cortex. Tumor resembles features of human high-grade glioma, including high cell density, mitoses, giant nuclei, and high density of Ki67 expression. White arrows indicate giant nuclei. c Confocal imaging series of tumor-bearing cortex on 7, 14, and 21 DPI (see “Methods”). Neurons indicated by NeuN (magenta), glial reaction indicated by GFAP (grey), and proliferating cells of growing tumor indicated by Ki67 (green). d Progression of tumor size on 7 DPI (green, N = 3 animals), 14 DPI (blue, N = 7 animals), and 21 DPI (magenta, N = 9 animals). (7 vs 14 DPI P = 0.7733; 14 vs 21 DPI P = 0.0067; 7 vs 21 DPI P = 0.0063) e Progressive changes of cell composition during tumor progression. Top row: Representative images from tumor-bearing cortical regions used for intensity analysis. Bottom row: fluorescent intensity analysis showing progressive reduction of neuron density (NeuN, magenta), expanding GFAP reactive rim (grey), and growing Ki67-positive tumor core (green) in time. Thin lines indicate individual animals (N = 2 animals per timepoint, see “Methods”), thick lines represent composite mean signal. The center of the tumor is indicated with dashed and the distance from tumor core is indicated on x-axis. f Confocal image of 21 DPI tumor-bearing cortex showing distinct borders of tumor core, infiltrated zone, and outer ridge. g Left: Example images of VGlut1/2 stainings (magenta), NeuN (cyan), Ki67 (green), and GFAP (blue) in peritumoral regions. Right: quantification of the densities of VGlut1/2 positive puncta per 25600 µm2 in the peritumoral regions. 7-DPI (N = 2, n = 9), 14-DPI (N = 2, n = 11), 21-DPI (N = 2, n = 10); peritumoral regions defined by contralateral (dark blue), outer ridge (grey), infiltrated zone (green), tumor core (light blue). DPI Days post injection. Error bars show mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. Statistics: d, g, one-way ANOVA with Sidak’s multiple comparisons test. Source data are provided as a Source Data file.
