Fig. 7: Manipulating AMPAR surface trafficking with R-mAbs.

a Approach for AMPAR ER retention. b Experimental timeline and images of cultured hippocampal neurons infected with AAVs encoding control (mNeon-KDEL) or designated i-mAb-KDEL, labeled with surface anti-GluA1 (cyan) or anti-GluA2 (magenta) (scale bars are 10 µm). c Quantification of GluA1 or GluA2 surface staining at different timepoints as a percentage of age-matched control neurons (n = 7 ROIs from 2 coverslips per timepoint). d mEPSC traces from cultured hippocampal neurons infected with designated AAVs (scale bar is 20 pA/5 s). e Quantification of mEPSC frequencies (mNeon-KDEL control n = 6, A1 scFv-mNeon-KDEL n = 9 neurons, two-tailed Student’s t-test, ****p < 0.0001; mNeon-KDEL control n = 9, A2 scFv-mNeon-KDEL n = 14 neurons, two-tailed Student’s t-test, ****p < 0.0001) and amplitudes (mNeon-KDEL control n = 6, A1 scFv-mNeon-KDEL n = 8, two-tailed Student’s t-test n.s. p = 0.7836; mNeon-KDEL control n = 9, A2 scFv-mNeon-KDEL n = 14 neurons, two-tailed Student’s t-test, *p = 0.0128). f Experimental setup and image of hippocampal slice expressing A1 scFv-mNeon-KDEL (scale bars: brightfield image 200 µm, fluorescence image 200 µm). g Control or A1 scFv-mNeon-KDEL expressing hippocampal slices were live labeled with A1 scFv-Halo/JF635i (magenta) prior to fixation and imaging (scale bar is 10 µm). Quantification (Relative Fluorescence Units) of A1 scFv-Halo/JF635i surface labeling is also plotted (control, n = 11 ROIs from 4 slices, A1 scFv-mNeon-KDEL n = 11 ROIs from 4 slices, two-tailed Student’s t-test ****p < 0.0001). h Example of a recorded (biocytin-filled) CA1 neuron to confirm A1 scFv-mNeon-KDEL expression (green) (scale bar is 10 µm). i Electrically-evoked currents were recorded at +40 mV (AMPAR + NMDARs) or −70 mV (AMPARs) from mNeon-KDEL control (black) or A1 scFv-mNeon-KDEL (red) neurons (scale bar is 100 pA/20 ms). Inset shows sEPSCs (scale bar is 20 pA/0.5 s). j AMPAR:NMDAR current ratios from mNeon-KDEL control or A1 scFv-mNeon-KDEL neurons (control n = 7 cells, A1 scFv-mNeon-KDEL n = 11 cells, two-tailed Student’s t-test, ****p < 0.0001). k sEPSC frequencies and amplitudes from mNeon-KDEL control or A1 scFv-mNeon-KDEL neurons (frequency: control n = 7 cells, A1 scFv-mNeon-KDEL n = 11 cells, two-tailed Student’s t-test, ****p < 0.0001) (amplitude: control n = 7 cells, A1 scFv-mNeon-KDEL n = 8 cells, two-tailed Student’s t-test, n.s. p = 0.7911). Data are presented as mean values +/- SEM. n.s. not significant. Source data are provided as a Source Data file.