Seizure evolution in a mouse model of West syndrome involves complex and time-dependent synapse remodeling, gliosis and alterations in lipid metabolism
Fig 6
Changes in morphology and lipid droplets of astrocytes in Cyfip2+/R87C mice.
(A) Representative 3D images of astrocytes 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 8 mice per genotype, two-way ANOVA with Šídák’s multiple comparisons test). (B) Serial electron microscopic images of lysosomes containing lipid droplets (LDs) in the hippocampal astrocytes of WT and Cyfip2+/R87C mice at PWK 28. 3D-rendered images of these lysosomes are also shown. (C) Correlative light and electron microscopy (CLEM) image of the hippocampal astrocyte in Cyfip2+/R87C mouse at PWK 28 shows BODIPY (light microscopy, LM), a marker for neutral lipids, within electron-dense lysosome observed by electron microscopy (EM). (D) Representative 3D images illustrate astrocytes (GFAP), lysosome (Lamp1) within the astrocytes, and LDs (BODIPY) within astrocytic lysosome (indicated by white arrows in merged images) in the hippocampus of WT and Cyfip2+/R87C mice at PWK 28, analyzed using Imaris software. Graphs show quantifications of the number and volume of LDs within astrocytic lysosomes in WT and Cyfip2+/R87C mice (n = 3 to 8 mice per genotype, unpaired two-tailed Student t test). (E) Serial electron microscopic images of a lipid droplet with crystals in the hippocampal astrocyte of Cyfip2+/R87C mouse at PWK 28. A 3D-rendered image of this lipid droplet is also presented. (F) CLEM image of the hippocampal astrocyte in Cyfip2+/R87C mouse at PWK 28 shows reflection signals from LM that match the crystals observed by EM. The graph displays the number of crystal spots, as measured by reflection signals from LM, in the hippocampus of WT and Cyfip2+/R87C mice at PWK 28 (n = 5 to 12 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.