Fig. 1

NMDAR KD in the recall-activated ensembles in the BLA and the mPFC. A Behavioral experiment scheme. 4-OHT was delivered two hours before remote retrieval 1, to tag ensembles activated during retrieval 1. B Left, a representative image of the recorded slice. Middle, sgGrin1 group showed a significantly lower level of NMDA/AMPA ratio than the sgGrin1 ATG group. Each dot represents a cell; grey, control group injected with scrambled sgRNA (n = 6 cells); orange, test group injected with sgGrin1 (n = 7 cells). Right, a representative EPSC trace of each group. Mann–Whitney test, *p = 0.0350. C Freezing level during retrieval 1. Unpaired t-test, ns, p = 0.7884. D Retention of fear memory after 4-OHT induction measured by the first tone block of the extinction curve. sgGrin1 ATG group, N = 13; sgGrin1 group, N = 11. Unpaired t-test, ns, p = 0.6485. E Trace of the freezing level during the extinction day 1 and 2. Each tone block represents the average freezing level of consecutive two tones. sgGrin1 ATG group, N = 13; sgGrin1 group, N = 11. Two-way repeated measures ANOVA; time effect, p = 0.0923; Grin1 KO effect, **p = 0.0068; F Extinction index of the sgGrin1 group and the sgGrin1 ATG groups. sgGrin1 ATG group, N = 13; sgGrin1 group, N = 11. Mann–Whitney test, *p = 0.0410. G Representative image of c-fos immunohistochemistry analysis. H Left panel, the proportion of tdT + neurons among DAPI + cells in the BLA. Each dot represents the average value of images from an individual mouse. sgGrin1 ATG group, N = 9; sgGrin1 group, N = 7; Mann–Whitney test, ns, p = 0.9182; right panel, Reactivation/chance = P(cfos|tdTomato)/P(cfos|DAPI); Mann–Whitney test, ns, p = 0.2105; One-sample Wilcoxon test, #p = 0.0156, ##p = 0.0039. I Left panel, the proportion of tdT + neurons among DAPI + cells in the mPFC. Each dot represents the average value of images from an individual mouse. sgGrin1 ATG group, N = 8; sgGrin1 group, N = 7; Mann–Whitney test, ns, p = 0.1893; right panel, Reactivation/chance = P(cfos|tdTomato)/P(cfos|DAPI); Mann–Whitney test, ns, p = 0.3357; One-sample Wilcoxon test, #p = 0.0156, ##p = 0.0078. Data are presented as mean ± SEM