Seizure evolution in a mouse model of West syndrome involves complex and time-dependent synapse remodeling, gliosis and alterations in lipid metabolism
Fig 5
Morphological and functional changes of microglia in Cyfip2+/R87C mice.
(A) Representative 3D images of microglia in the hippocampal CA1 region of WT and Cyfip2+/R87C mice at PWK 14 and 28, processed using Imaris software. Graphs show quantifications from Sholl analysis as well as measurements of branch length, area, points, and diameter (n = 7 to 9 mice per genotype, two-way ANOVA with Šídák’s multiple comparisons test). (B) Representative fluorescence immunohistochemistry images and quantification show increased cleaved Caspase3 signals in microglia of the Cyfip2+/R87C hippocampus compared to the WT hippocampus at PWK 28, but not at PWK 14 (n = 7 to 9 mice per genotype, two-way ANOVA with Šídák’s multiple comparisons test). Arrows indicate microglia with enlarged images shown in the insets. SO, stratum oriens, SP, stratum pyramidale, SR, stratum radiatum. (C) Representative fluorescence immunohistochemistry images and quantification show increased CD68 signals in microglia of the Cyfip2+/R87C hippocampus compared to the WT hippocampus at PWK 14 (n = 7 to 8 mice per genotype, unpaired two-tailed Student t test). (D) Heat maps display qRT-PCR results for genes related to microglial activation in the cortex and hippocampus of Cyfip2+/R87C mice compared to WT mice at different ages (n = 6 mice per genotype, two-way ANOVA with Šídák’s multiple comparisons test). Statistical significance of expression differences for each gene between PWK 14 and 28 within Cyfip2+/R87C mice is shown in the ‘PWK 14 vs. 28’ column. Note that Il-6 was not detected in the cortex at PWK 1. (E) Representative 3D images illustrate microglia (IBA1), lysosome (Lamp1) within the microglia, and excitatory synaptic protein (Homer1) within microglial lysosome (i.e., engulfed Homer1, indicated by white arrows in merged images) in the hippocampal CA1 region of WT and Cyfip2+/R87C mice at PWK 14, analyzed using Imaris software. Graphs show quantifications of microglial lysosome and engulfed Homer1 in WT and Cyfip2+/R87C mice (n = 7 mice per genotype, unpaired two-tailed Student t test). *P < 0.05; **P < 0.01; ***P < 0.001. Data are represented as mean ± standard error of the mean. The data underlying this Figure can be found in S1 Data.