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. 2023 Sep 27;9(39):eadg8148. doi: 10.1126/sciadv.adg8148

Fig. 3. The effects of immune-mediated astroglial activation on hippocampal neurogenesis and cognitive performance.

Fig. 3.

(A) Schematic diagram of assays for hippocampal neurogenesis (EdU and BrdU labeling) and cognitive performance with stereotaxic injections of control (msCtrl-IgG) or monoclonal mouse anti-AQP4 antibodies (msAQP4-IgG). (B) Analysis of AQP4 expression in astrocytes [at end-feet along microvessels (CD31+)] in msCtrl-IgG– or msAQP4-IgG–injected hippocampus. n = 3 animals. Scale bar, 20 μm. (C) Quantification of astrogliosis (GFAP) by msAQP4-IgG treatment within the volume of injected DG determined by 3D rendering of confocal images. n = 3. Scale bar, 100 μm. (D) Assessment of CHI3L1 expression in astrocytes induced by msAQP4-IgG. n = 3. Scale bar, 20 μm. (E) Representative movement paths of msCtrl-IgG– or msAQP4-IgG–injected mice in MWM. (F) MWM platform crossing numbers. n = 9. (G) Escape latencies to find the platform in MWM. n = 9. (H) Representative images and analyses of the proliferating cells (EdU+) in hippocampus SGZ. n = 5. Scale bar, 100 μm. (I) Quantification of radial glia–like NSCs (EdU+GFAP+Sox2+) and transiently amplifying progenitor-like cells (EdU+GFAP−Sox2+). n = 4. Scale bar, 100 μm. (J) Analysis of immature neurons (DCX+) differentiated from NSCs (BrdU+) after msCtrl-IgG or msAQP4-IgG injections. n = 4. Scale bar, 100 μm. (K) Analysis of mature neurons (NeuN+) differentiated from NSCs (BrdU+) after msCtrl-IgG or msAQP4-IgG injections. n = 4. Scale bar, 100 μm. (L) Schematic diagram of assays for hippocampal neurogenesis and cognitive performance with mini-pump infusion of msCtrl-IgG or msAQP4-IgG. (M to O) Quantification of total proliferating (EdU+) cells, radial glia–like NSCs (EdU+GFAP+Sox2+), and transiently amplifying progenitor-like cells (EdU+GFAP−Sox2+) in IgG-infused DG. n = 4. (P and Q) Quantification of newborn immature (BrdU+DCX+) neurons and mature (BrdU+NeuN+) neurons in IgG-infused DG. n = 4. Bar graphs presented in means ± SEM; (B) to (D), (F), (H) to (K), and (M) to (Q), by Student’s t test; (G), by two-way ANOVA with Tukey’s multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.001.