Extended Data Figure 6: Phase-specific optogenetic inhibition of dmPFC PNs. | Nature

Extended Data Figure 6: Phase-specific optogenetic inhibition of dmPFC PNs.

From: Prefrontal neuronal assemblies temporally control fear behaviour

Extended Data Figure 6

a, Twenty-four hours after conditioning, levels of freezing in wild type (n = 7) and GFP (n = 7) control animals where similar (2 min baseline, four CS+ and four post-CS+ periods of 30 s averaged, two-way ANOVA, factor 1: wild type versus GFP, P > 0.05; factor 2: time, P < 0.001; F1 × F2, P < 0.01, Student–Newman–Keuls post-hoc test within wild type and GFP, ***P < 0.001). b, Corresponding time-resolved changes in freezing behaviour around CS+ offset for wild type and GFP control animals (two-way ANOVA, factor 1: wild type versus GFP, P > 0.05; factor 2: time, P > 0.05; F1 × F2, P > 0.05). c, Comparison of GFP control levels of freezing during (CS+) and 30 s after CS+ (post-CS+) on days 2 (n = 7), 3 (n = 7) and 4 (n = 4) (2 min baseline, four CS+ and four post-CS+ periods of 30 s averaged; two-way ANOVA, factor 1: day, P > 0.05; factor 2: time, P < 0.001; F1 × F2, P > 0.05; Bonferroni post-hoc test between epochs, **P < 0.01, ***P < 0.001). d, Timeline scheme for the optogenetic stimulation protocol on day 2 with stimulation targeting the ascending phase of 4 Hz oscillation cycle (top) and day 3 with stimulation targeting the descending phase of 4 Hz oscillation cycle (bottom). e, f, Comparison of freezing levels between CS+ 1 and CS+ 2–4 for epochs 10 s preceding, during and 30 s after CS+ during optogenetic stimulation (pre-CS+, CS+ and post-CS+ respectively) on day 2 (e) and day 3 (f). On day 2, ascending phase stimulation after CS+ 1 induced a decrease of freezing levels during subsequent CS+ epochs as no more difference was observed between pre CS+, CS+ and post CS+ for CS+ 2–4 compared with CS+ 1 (n = 7 mice, repeated-measures two-way ANOVA, factor 1: CS+ 1 versus CS+ 2–4, P > 0.05; factor 2: epoch, P < 0.05; F1 × F2, P > 0.05; Student–Newman–Keuls post-hoc test within factor 1, *P < 0.05 compared with both pre- and post-CS+ epochs). On day 3, descending phase stimulation after CS+ 1 induced an increase in freezing levels during pre-CS+ epochs as no more difference was observed between pre-CS+, CS+ and post-CS+ for CS+ 2–4 compared with CS+ 1 (n = 7 mice, repeated-measures two-way ANOVA, factor 1: CS+ 1 versus CS+ 2–4, P > 0.05; factor 2: epoch, P < 0.05; F1 × F2, P < 0.05; Student–Newman–Keuls post-hoc test within factor 1, *P < 0.001 compared with both CS+ and post-CS+ epochs). g, Averaged freezing behaviour evoked by CS+ 1 presentation on day 3 if day 2 stimulation was in the ascending phase (GFP n = 7, ChR2 with day 2 ascending n = 7, Mann–Whitney rank sum test, P = 0.259).

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