Article Figures & Data
Figures
- Figure 1.
Aβ-induced synaptic alterations coincide with enhanced AMPAR ubiquitination. A, Representative traces of mEPSCs from each condition indicated. Calibration: 500 ms, 20 pA. B, C, Mean mEPSC amplitude (B) and frequency (C) for neurons (DIV 16–20) treated with CHO or 7PA2 CM for 3 or 6 d. D, Mean mEPSC amplitude for neurons treated with DMSO or oligomeric Aβ for 3 d. E, Representative immunofluorescent images of dendrites treated with CHO or 7PA2 CM for 6 d. Scale bar, 5 μm. F, Quantification of spine density from conditions displayed in E. G, Representative Western blot of total and biotinylated surface GluA1 in neurons treated with CHO or 7PA2 CM for 3 d. H, Quantification of immunoblots for total and surface GluA1, normalized to CHO. I, Representative immunoblot depicting enhanced AMPA-induced (100 μm, 10 min) ubiquitination of AMPARs following a 14 h treatment with 7PA2 CM. *p < 0.05. **p < 0.01. ***p < 0.001.
- Figure 2.
Aβ promotes Nedd4-1 redistribution into dendritic spines and increases Nedd4-1 association with surface proteins. A, Representative immunofluorescent images of hippocampal dendrites expressing HA-tagged Nedd4-1, treated with CHO or 7PA2 CM for 23 h and stained with anti-GFP (green) and anti-HA (white) antibodies. Scale bar, 5 μm. B, Quantification of HA-Nedd4-1 fluorescence local maxima from conditions displayed in A. C, Normalized ratio of spine to total HA levels between conditions in A. D, Representative Western blot of surface and total Nedd4-1 and GluA1 in neurons treated with CHO or 7PA2 CM for 3 d. E, Quantification of immunoblots for surface-associated Nedd4-1, normalized to CHO. *p < 0.05. **p < 0.01. ***p < 0.001.
- Figure 3.
Nedd4-1 is necessary for Aβ-induced synaptic alterations. A, Representative traces of mEPSCs from each condition indicated. Calibration: 500 ms, 20 pA. B, Cumulative probability distributions of all mEPSC amplitudes recorded from each condition. Inset, Averaged trace for all mEPSCs in each group; n = 1960, n = 2287, n = 1468, n = 2302 events. Calibration: 5 ms, 5 pA. C, Mean mEPSC amplitude for cells in all conditions. D, Representative Western blot of surface GluA1 levels in neurons treated with CHO or 7PA2 CM. E, Quantification of immunoblots for surface GluA1, normalized to CHO within each virus. F, Representative immunofluorescent images of dendritic spines from hippocampal neurons under each of the conditions indicated. Scale bar, 5 μm. G, Quantification of spine density from conditions displayed in F. ***p < 0.001.
