Fig. 3
NMDAR-Ab from Healthy + subjects and PSY + patients display different effects on synaptic NMDAR dynamics. a Schematic representation of the experimental design. b Representative trajectories of a single GluN2A-NMDAR-QD complex (500 frames, 50 ms acquisition) exposed for 30 min to purified IgG (5 µg ml−1) from Healthy + or PSY + subjects within synaptic areas. Scale bar, 500 nm. c Comparison of GluN2A-NMDAR instantaneous diffusion coefficient (µm2 s−1) within the postsynaptic density (PSD) area in the presence of a commercial αGluN1N-term antibody (n = 749 trajectories, N = 52 neurons) or purified IgG from Healthy − (n = 94, N = 13), Healthy + (n = 313, N = 42) or PSY + (n = 695, N = 59) subjects (***P < 0.0001, Kruskal–Wallis followed by Dunn’s multiple comparison test). Data are expressed as median diffusion coefficient ± 25–75% IQR. d Mean square displacement (MSD) over time of GluN2A-NMDAR within the PSD area after exposure to a commercial αGluN1N-term antibody or purified IgG from Healthy −, Healthy +, and PSY + individuals (P < 0.001, Kolmogorov–Smirnov test). Data are expressed as mean ± SEM (dash lines). e Mean ± SEM of diffusive GluN2A-NMDAR exchange frequency between synaptic and extrasynaptic compartments in Healthy + (n = 241 trajectories) or PSY + (n = 268) conditions (***P < 0.001, Mann–Whitney test). f MSD over time of GluN2A-NMDAR within the perisynaptic area exposure to Healthy + or PSY + NMDAR-Ab for 30 min (P = 0.076, Kolmogorov–Smirnov test). Data are expressed as mean ± SEM (dash lines). Inset: GluN2A-NMDAR instantaneous diffusion coefficient within the perisynaptic area in Healthy + (n = 1344 trajectories, N = 42 neurons) or PSY + (n = 2264, N = 59) conditions (P = 0.213, Mann–Whitney test). g Comparison of GluN2A-NMDAR instantaneous diffusion coefficient after 30 min exposition to various concentrations of NMDAR-Ab. Data are expressed as median diffusion coefficient ± 25-75% IQR. Healthy + 50 ng ml−1, n = 321 trajectories, N = 21 neurons; Healthy + 5 µg ml−1, n = 69, N = 32; PSY + 50 ng ml−1, n = 187, N = 24; PSY + 5 µg ml−1, n = 178, N = 32. *P < 0.05, ***P < 0.001, Kruskal–Wallis followed by Dunn’s multiple comparison test. h Experimental design of the NMDAR-Ab pre-absorption experiment. Right panel: normalized GluN2A instantaneous diffusion coefficient in control (n = 1,388 trajectories, N = 37 neurons), PSY + (n = 2,658, N = 59), NT-HEK (n = 3867, N = 27), and GluN1-HEK (n = 4799, N = 28) conditions (***P < 0.001, Kruskal–Wallis followed by Dunn’s multiple comparison test)
