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. Author manuscript; available in PMC: 2019 May 31.
Published in final edited form as: Cell. 2018 May 3;173(6):1329–1342.e18. doi: 10.1016/j.cell.2018.04.004

Figure 6. Photoinhibition of ACC→BLA impairs observational fear conditioning, but not classical fear conditioning.

Figure 6

A. Viral injection and optic fiber placement for selective inhibition of ACC→BLA circuit.

B. Behavioral and light delivery paradigm for inhibition of ACC→BLA circuit during cue presentations during acquisition (Day 1) of observational conditioning.

C. During observational conditioning, there were no significant differences in freezing between NpHR (N=7) mice and eYFP (N=12) mice (unpaired, two-tailed t-test, t=0.0785, df=17, P =0.9383). However, on Test day, cue driven freezing was impaired in NpHR compared to eYFP mice (unpaired, two-tailed, t-test, t=2.378, df=17, *P =0.0294). Insets show cue and baseline (20s prior to cue onset) freezing values for observational conditioning and test day (BL=baseline; Observational conditioning: two-way ANOVA, group effect, F(1,17)=8.286, P=0.0104, epoch effect, F(1,17)=66.26, P<0.0001, group X epoch interaction, F(1,17)= 0.0829, P0.7769; Bonferroni post-hoc analysis, ****P< 0.0001, ***P=0.0002; Test day: two-way ANOVA, group effect, F(1,17)= 0.3596, P=0.5566, epoch effect, F(1,17)=10.64, P=0.0046, group X epoch interaction, F(1,17)=5.657, P=0.0294; Bonferroni post-hoc analysis, ***P=0.0005).

D. Behavioral and light delivery paradigm for inhibition of ACC→BLA circuit during cue presentations during expression (Day 2) of observational conditioning.

E. There were no significant differences in cue driven freezing between NpHR (N=9) and eYFP (N=8) mice during observational conditioning (unpaired, two-tailed, t-test, t=0.4916, df=15, *P =0.6301) or Day2: Test (unpaired, two-tailed, t-test, t=0.5137, df=15, *P =0.6149). Insets show cue and baseline (20s prior to cue) freezing values during conditioning and test day (Observational conditioning: two-way ANOVA, group effect, F(1,15)=10.46, P=0.0056, epoch effect, F(1,15)=18.17, P=0.0007, group X epoch interaction, F(1,15)=0.2416, P=0.6301; Bonferroni post-hoc analysis, *P<0.05; Test day: two-way ANOVA, group effect, F(1,15)= 12.30, P=0.0032, epoch effect, F(1,15)=6.778, P =0.02, group X epoch interaction, F(1,15)=0.06837, P=0.7973; no significant Bonferroni post-hoc analysis).

F. Inhibition of ACC→BLA circuit during classical fear conditioning. No significant differences were detected between NpHR (N=7 mice) and eYFP (N=10) mice in cue driven freezing on test day (unpaired, two-tailed, t-test, t=1.02, df=15, *P=0.3237). Inset shows cue and baseline (20s prior to cue) freezing values (two-way ANOVA, group effect, F(1,15)=0.0061, P =0.9389, epoch effect, F(1,15)= 28.48, P<0.0001, group X epoch interaction, F(1,15)=1.041, P=0.3237; Bonferroni post-hoc analysis, **P<0.01). All error bars indicate ± SEM.