Fig. 5: Restoration of mucin-type O-glycosylation increases neuronal homeostasis, reduces glial inflammation and improves memory and learning in aged mice.
From: Glycocalyx dysregulation impairs blood–brain barrier in ageing and disease
a, Experimental scheme used for behavioural testing of mice. b, Spatial working memory assessment using the Y maze (n = 26 (aged EGFP), 19 (C1GALT1), 23 (B3GNT3) and 20 (young EGFP); one-way ANOVA with Tukey’s post hoc test; mean ± s.e.m.). c, Hippocampal-dependent learning and memory assessment by contextual fear conditioning (n = 26 (EGFP), 18 (C1GALT1) and 19 (B3GNT3); one-way ANOVA with Dunnett’s post hoc test; mean ± s.e.m.). d, Outline of snRNA-seq profiling of pooled cortical and hippocampal tissue of young AAV-EGFP, aged AAV-EGFP and aged AAV-B3GNT3-treated groups. e, Uniform manifold approximation and projection (UMAP) of 69,250 nuclei from pooled cortical and hippocampal tissue of young AAV-EGFP, aged AAV-EGFP and aged AAV-B3GNT3-treated groups (n = 3 mice per group), coloured by cell type. BEC, brain endothelial cell; ExN, excitatory neuron; InN, inhibitory neuron; MG, microglia; OPC, oligodendrocyte precursor cell. f, Overview of key DEGs in each major cell type induced by AAV-B3GNT3 treatment in aged mice compared with AAV-EGFP. Astro, astrocyte. g, Volcano plot of ExN DEGs induced by AAV-B3GNT3 treatment in aged mice (upregulated genes in pink and downregulated genes in red). Padj, adjusted P value. h, Comparison of ExN DEG fold changes with B3GNT3 overexpression (y axis) and reverse ageing (x axis). Areas in which AAV-B3GNT3 treatment causes changes in the reverse direction of ageing are highlighted. i, Top upregulated pathways in ExN based on DEGs shared between AAV-B3GNT3 treatment and reverse ageing (top right quadrant in h). j, IBA1 and CD68 expression in the cortices of AAV-transduced mice. Scale bars, 50 µm. k, Quantification of CD68+ signal in IBA1+ microglia in j (n = 5 mice per group; one-way ANOVA with Tukey’s post hoc test; mean ± s.e.m.). l, Quantification of cortical IBA1+ area in j (n = 5 mice per group; one-way ANOVA with Tukey’s post hoc test; mean ± s.e.m.).
