Plasma membrane remodeling in GM2 gangliosidoses drives synaptic dysfunction
Fig 5
A model of GM1 Gangliosidosis shares lysosomal proteomic changes and hallmarks of lysosomal exocytosis.
A. Quantitative PCR (qPCR) analysis of GLB1 gene expression in neurons following CRISPRi-induced knockdown at 14 dpi. Fold change relative to SCRM controls are shown for N = 3 biological replicates carried out in technical triplicate n = 3 (squares, triangles, circles) and the mean is displayed (red line). Significance was determined with an unpaired t test, ****p ≤ 0.0001. B. Quantification of whole-cell gangliosides at 28 dpi for SCRM, ΔGLB1 cells, the mean of N = 3 biological replicates is displayed ± SEM. Significance was determined by two-way ANOVA, ****p ≤ 0.0001. C. Analysis of quantitative WCP data at 28 dpi for ΔHEXA neurons compared with SCRM cells vs. ΔGLB1 neurons compared with SCRM cells. Fold change in protein abundance for ΔHEXA cells (x-axis) are plotted against fold change in protein abundance for ΔGLB1 cells (y-axis). Significance (2-sided t test across N = 3 biological replicates) is illustrated as those significantly changed in ΔGLB1 only (blue), ΔHEXA only (red) or in both (purple, large data points). D. Select shared targets from the WCP when both gangliosidosis models are combined are represented graphically to illustrate the fold change in whole cell protein abundance. E. GO term analysis for proteins significantly changed in the WCP for both gangliosidosis models is shown for cellular component, molecular function and biological process with the change plotted as the false discovery rate (log10FDR) and the number of proteins in each group indicated. F. Quantitative mass spectrometry following enrichment of PM proteins from ΔHEXA and ΔGLB1 gangliosidosis models compared with the SCRM control. A volcano plot is shown with average fold change (x-axis) and significance (y-axis, two-sided t test) across N = 3 biological replicates for each of the ΔHEXA and ΔGLB1 lines compared to N = 2 SCRM. Proteins that are significantly changed (p ≤ 0.05) are colored (red). Underlying data used to generate these figures are available in S1 Data at https://doi.org/10.17863/CAM.118836.