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. 2023 Mar 15;133(6):e164919. doi: 10.1172/JCI164919

Figure 7. Grn–/– astrocytes promote synaptic and dendritic degeneration.

Figure 7

(A and B). IHC and confocal images of GJA1 (A) and EAAT2 (B) in astrocytes in cortex (CX) and thalamus (TH) in 19-month-old Grn+/+ and Grn–/– mice. Scale bars: 1 mm; 100 μm (inset) (C–E) Confocal images of PSD-95, Bassoon, and GFAP in mTH (C) and layer 1/2 (L1/2) (D) and layer 6 (L6) (E) in mFCX in 19-month-old Grn+/+ and Grn–/– mice. Scale bars: 10 μm; 5 μm (inset). (F) Quantification of PSD-95+;Bassoon+ area within GFAP+ astrocytes in Grn+/+ and Grn–/– mice (n = 3 each). Statistics uses 2-tailed Student’s t test. (G) Schematic diagram for astrocyte-neuron cocultures. (H) Confocal images of MAP2, SYP, and GFAP in astrocyte-neuron cocultures. Scale bars: 50 μm and 20 μm (inset). (I) Quantification of cumulative SYP+ (left) and PSD95+ (right) density in Grn+/+ and Grn–/– dendrites near Grn+/+ or Grn–/– astrocytes. Data from 2 independent cocultures. Statistics uses 2-way ANOVA. (J) Confocal images of MAP2+ neurons and GFAP+ astrocytes in astrocyte-neuron cocultures. Scale bars: 50 μm. (K) Quantification of MAP2+ dendritic arborization in Grn+/+ or Grn–/– neurons cocultured with Grn+/+ or Grn–/– astrocytes. Data from 4 independent cocultures. Statistics uses 2-way ANOVA. (L) Confocal images of Bassoon, PSD-95, and MAP2 in Grn+/+ and Grn–/– cortical neurons treated with control media, Grn+/+ ACM, or Grn–/– ACM. Insets are higher magnification images from boxed areas above. (M and N) Quantification of Bassoon+;PSD-95+ synaptic density in the dendrites of Grn+/+ and Grn–/– neurons (M) and cleaved caspase 3+ Grn+/+ and Grn–/– neurons (N) treated with control media, Grn+/+ ACM, or Grn–/– ACM. Data are from 4 independent cocultures. 1-way ANOVA. All quantification data represent mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.0001. NS, not significant.