Fig. 2: Optogenetic manipulation of ACh signal in the ACx at the encoding stage regulates the DAFM. | Translational Psychiatry

Fig. 2: Optogenetic manipulation of ACh signal in the ACx at the encoding stage regulates the DAFM.

From: ACx-projecting cholinergic neurons in the NB influence the BLA ensembles to modulate the discrimination of auditory fear memory

Fig. 2

A Left, schematic of virus injection. In ChAT-cre mice, AAV9-DIO-NpHR3.0 or AAV9-DIO-ChrimsonR-mcherry was infused into the NB; AAV9-Syn-ACh3.0 was infused into the ACx. Right: Confocal images of coronal sections showing Cre+ neurons (mcherry+) in the NB of ChAT-cre mice were co-labeled with cholinergic neurons (green) Scale bar: 200 μm; 20 μm. B Behavioral design. C, E Percentage of the freezing time (left, middle) and discrimination index (right) in mice under inhibition (C) or activation (E) of ACx-projecting cholinergic neurons of the NB at the encoding stage (C, NpHR3.0, n = 10, mCherry, n = 10; F (1, 36) = 13.96, P < 0.001; E ChrimsonR, n = 8; mCherry, n = 8; F (1, 28) = 0.0059, P = 0.94). D, F Left: The mean and SEM of the fluorescence responses of ACh3.0 in the ACx of mice at day 2 receiving inhibition (D) or activation (F) of the ACx-projecting cholinergic neurons of the NB at the encoding stage. Red line indicated bins between the 10k and 1k trials with two-tailed unpaired t-test significance (P < 0.05). Middle and Right: quantification of the average amplitude of increased signal against baseline between control (Fig. 1D, right) and NpHR3.0 (Fig. 2D, left) or ChrimsonR (Fig. 2F, left) group during the 1 kHz tone retrieval (middle) or 10 kHz tone retrieval (right). G Schematic of virus injection of AAV9-DIO-NpHR3.0-mcherry in the NB and behavioral design. H Percentage of the freezing time (left, middle) and discrimination index (right) in mice under either or both nicotine intraperitoneal injection and inhibition of ACx-projecting cholinergic neurons of NB at the encoding stage (mCherry-saline, n = 8; mCherry-nicotine, n = 9; NpHR3.0-saline, n = 11; NpHR3.0-nicotine, n = 11; freezing: F (3, 70) = 7.862, P < 0.001; discrimination index: F (3, 35) = 21.02, P < 0.001). Statistical tests: C, E, and H, freezing: two-way ANOVA with Bonferroni’s multiple comparisons tests. C, E discrimination index: unpaired two-tailed t-test. H discrimination index: one-way ANOVA with Bonferroni’s multiple comparisons tests. D, F signal variation: unpaired two-tailed Mann–Whitney-U-test. ns not significant; *P < 0.05, **P < 0.01, ***P < 0.001. Data are presented as mean ± SEM.

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