Fig. 5: Selective inhibition of the activity of glutamatergic TANsACC neurons weakened the positive effects of taVNS on PTSD-related anxiety-like behaviors.
(A) Schematic of AAV-CaMKIIα-DIO-hM4Di-mCherry or AAV-CaMKIIα-DIO-mCherry injection into the ACC of Fos-TRAP2 mice (Created with BioRender.com, 2025). (B) Representative images showing the infusion site within the ACC (left) and mCherrylabeled neurons co-expressed with glutamate (Glu) antibody (right), scale bar, 250 μm (left), and 75 μm (right). (C) Representative trace (left) and summarized data (right) showing bath application of CNO (10 μM) hyperpolarizes the glutamatergic TANsACC neurons (n = 7 neurons from 3 mice). (D) Representative traces during OFT in mCherry and hM4Di mice. (E-G) Entry times (E), duration (F), and traveled distance (G) in the central area of the open field of two groups [Two-tailed unpaired t-test: (E) t(21) = 4.2590, p = 0.0003. (F) t(21) = 3.2700, p = 0.0037. (G) t(21) = 4.480, p = 0.0002]. (H) Representative traces during EPMT in two groups. (I-K) Entry times (I), and duration (J) in the open arms, as well as anxiety index (K) of two groups [Two-tailed unpaired t-test: (I) t(21) = 3.841, p = 0.0010. (K) t(21) = 3.6550, p = 0.0015. Mann-Whitney U test: (J) Z = −2.9860, p < 0.0001]. (L) Representative traces during TCT in two groups. (M, N) Sniffing time (M), and social index (N) of two groups [(M) Two-tailed unpaired t-test: mCherry: E vs S1, t(18) = 8.1850, p < 0.0001; Mann-Whitney U test: hM4Di: E vs S1, Z = −4.3333, p < 0.0001. (N) Two-tailed unpaired t-test: t(21) = 1.020, p = 0.3193]. mCherry: n = 10 mice, hM4Di: n = 13. Data are represented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
