Fig. 4

Treatment with riluzole prevents cranial radiation-induced reduction in the neuronal plasticity-related cFos expression. A, a1 Dual immunofluorescence staining, confocal microscopy and 3D algorithm-based unbiased quantification of NeuN⁺ (green) neuronal plasticity-related immediate early gene (IEG) expression product, cFos showed a significant reduction in the number of cFos-NeuN⁺ neurons in the irradiated (RT + Vehicle) hippocampal sub-granular zone (SGZ) and the granule cell layer (GCL) compared to Control + Vehicle group. B–E Single channel fluorescent z stacks for NeuN (green), cFos (red) and merged images show cFos-NeuN co-expression in the hippocampal dentate gyrus for each treatment group. a1 Higher magnification 3D fluorescent z stack of dual-stained neuron (arrow) is shown for the Control + Vehicle group B. Treatment with riluzole significantly increased the number of cFos-NeuN⁺ neurons in the irradiated hippocampus (RT + RZ) compared to irradiated mice receiving vehicle (RT + Vehicle). Data is presented as mean ± SEM (N = 8 mice per group). P values were derived from two-way ANOVA and Bonferroni's multiple comparisons test. Scale bars, 100 μm, (B–E), and 5 μm, (a1)