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. 2018 Jul 23;25(11):2832–2843. doi: 10.1038/s41380-018-0124-3

Fig. 2.

Fig. 2

(a, b) NMDAR/AMPAR ratio values from neurons in cont (gray circles; n = 8 cells, 5 mice) and DG-GluN1 KO mice (orange circles; n = 7 cells, 6 mice). Hash marks indicate mean values ratio in cont and DG-GluN1 KO. Both biophysical (a) and pharmacological (b) approaches showed that NMDAR/AMPAR ratio in cont is similar to that of found in DG-GluN1 KO mice (p > 0.05). For biophysical approach, AMPAR- and NMDAR-EPSC amplitudes are extracted from the dual component obtained at +40 mV, at 10 and 50 ms post-stimulus, respectively. For pharmacological approach: D-APV at 50 μM was used to extract AMPAR-mediated current. (b) Right panel: sample EPSCs from cont (WT) and DG-GluN1 KO mice. Calibration: 50 ms, 20 pA. (c, d) Both AMPAR- (c) and NMDAR-mediated transmission (d) at MF-CA3 synapses are increased. AMPAR-mediated transmission was assessed at −70 mV, and NMDAR-mediated current was elicited at +40 mV. Right panels for (c) and (d): example of AMPAR- (calibration: 10 ms, 50 pA) and NMDAR-EPSCs traces (calibration: 50 ms, 20 pA) from a cont and a DG-GluN1 KO neuron over the stimulus range 0, 20, 40, 60, 80, and 100 μA. Cont, n = 10 cells, 4 mice; DG-GluN1 KO, n = 16 cells, 4 mice. Two-way ANOVA: genotype and genotype × stimulation interaction effects for both AMPAR- (genotype effect: F(1, 24) = 4.312, *p = 0.048, and interaction effect: F(4, 96) = 3.850, **p = 0.006) and NMDAR-EPSCs (genotype effect: F(1, 17) = 5.336, *p = 0.033, and interaction effect: F(8,136) = 6.160, ***p < 0.0001). (e) Left panel, percentage of cells demonstrating eplileptiform activity indicated by more frequent late burst EPSC recruitment achieved at lower stimulus intensities was significantly higher at DG-GluN1 KO mice (12/16 cells) compared with cont (2/10 cells). Right panel, sample traces from CA3 pyramidal neurons from DG-GluN1 KO mice. Calibration in left: 20 ms, 50 pA; calibration in right: 100 ms, 1 nA. (f) Left, examples of evoked dual EPSCs at membrane potentials from −80 to +40 mV. Calibration: 10 ms, 50 pA. Right, IV relationship for AMPAR-EPSCs (measured by extracting the AMPAR current from the dual component at 10 ms post stimulus) in cont and DG-GluN1 KO mice (n = 8–10 cells, 5 mice in each group). The lines represent the linear regression (r = 0.99 for each group). (g) Left, examples of evoked NMDAR-mediated EPSCs at membrane potentials from −80 to +40 mV. Calibration: 50 ms, 20 pA. Right, IV relationship for NMDAR-mediated EPSCs in cont and DG-GluN1 KO mice (n = 5–11 cells, 3–4 mice in each group). Holding potentials were not corrected for liquid junction potential. Data are represented as means ± SEM