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

Figure 8. iPSC-derived PGRN-deficient astrocytes fail to promote synaptic refinement, but enhance neuronal stress and TDP-43 proteinopathy in cortical organoids.

Figure 8

(A) Schematic diagram for transplanting GRN+/+ and GRN–/– iPSC-derived astrocytes (iAST) to organoids. (B) Confocal images of GFP+ GRN+/+ iAST and GRN–/– iAST in organoids 1-week after transplantation. Scale bars: 500 μm. (C) Confocal and IMARIS images of GFP, PSD-95, and Bassoon in GRN+/+ iAST and GRN–/– iAST within the organoids. Scale bars: 10 μm and 3 μm (inset and IMARIS). (D) Quantification of PSD-95 puncta size outside and inside GFP+ area in organoids transplanted with GRN+/+ iAST (n = 10), GRN–/– iAST (n = 9), or no iAST (n = 4). Statistics use 1-way ANOVA (upper panel) and 2-tailed Student’s t test (lower panel). (E) Confocal and IMARIS images of GFP and GJA1 in GRN+/+ iAST and GRN–/– iAST in organoids. Scale bars: 20 μm. (F) Quantification of GJA1+ surface area in organoids with GRN+/+ iAST (n = 10) or GRN–/– iAST (n = 9), Statistics use 2-tailed Student’s t test. (G) Confocal and IMARIS images of GFP and EAAT2 in GRN+/+ iAST and GRN–/– iAST in organoids. Scale bars: 20 μm. (H) Quantification of EAAT2+ area in GFP+ GRN+/+ iAST (n = 10) and GFP+ GRN–/– iAST (n = 9). Statistics use 2-tailed Student’s t test. (I) Confocal images of GFP and PGK1 in organoids transplanted with GRN+/+ or GRN–/– iASTs. Scale bars: 20 μm. (J) Quantification of PGK1+ area in MAP2+ neurons in organoids transplanted with GRN+/+ iASTs (n = 12) or GRN–/– iAST (n = 10). Statistics use 2-tailed Student’s t test. (K) Confocal and IMARIS images of TDP-43 and NeuN in GRN+/+ iAST and GRN–/– iAST-containing organoids. Scale bars: 10 μm and 5 μm (inset and IMARIS) (L) Quantification of TDP-43+ signals in extranuclear (enTDP-43) area (top) and particle size (bottom) in organoids transplanted with GRN+/+ iAST (n = 11), GRN–/– iAST (n = 10), or no iAST (n = 4). 1-way ANOVA. All quantification data represent mean ± SEM, and n represents the number of independent samples tested. *P < 0.05, **P < 0.01.