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. 2024 Oct 29;7:1408. doi: 10.1038/s42003-024-07105-5

Fig. 7. Activating GABAergic interneurons in the BLA by chemogenetic regulation attenuated the effect of propofol on enhancing fear memory.

Fig. 7

a Experimental time course for surgery, CNO injection, FC training, propofol injection, and FCT. b Schematic of 2/9AAV-DIO-hM3D(Gq)-mCherry or 2/9AAV-DIO-mCherry and rAAV-CaMKIIa-EYFP injection into the BLA of Vgat-cre mice. Representative immunohistochemical staining (scale bar, 1000 µm, AP: -1.05 mm, ML: ±3.15 mm, DV: 4.8 mm). c Neurons expressing mCherry fluorescence were observed in the BLA when in vitro brain slices were exposed under a microscope. Bath application of CNO (10 µM) increased the firing rate in hM3Dq-positive BLA neurons in vitro. d Quantification of the firing rate (n = 14 cells from 6 mice injected with AAV-VEH and n = 14 cells from 6 mice injected with AAV-hM3Dq, two-tailed Wilcoxon matched-pairs signed rank test, *p = 0.013). e Chemogenetic activation was applied to the BLA slice, and the activity of glutamatergic neurons was recorded. Bath application of CNO (10 µM) decreased the firing rate in BLA glutamatergic neurons in vitro. f Quantification of the firing rate (n = 12 cells from 6 mice injected with AAV-VEH+ rAAV-CaMKIIa-EYFP and n = 13 cells from 6 mice injected with AAV-hM3Dq+ rAAV-CaMKIIa-EYFP, two-tailed paired t test, **p < 0.01). g Representative Western blot image and quantification of c-Fos expression. The expression of c-Fos was reduced after chemogenetic activation of GABAergic interneurons in the BLA (n = 8, two-tailed unpaired t test, *p = 0.045). h Representative images of mCherry/GAD67/c-Fos immunofluorescence in BLA neurons after virus treatment and chemogenetic manipulation; scale bar, 100 µm. I–k The ratio of c-Fos+ & GAD67+ cells in the BLA was increased after chemogenetic activation of GABAergic interneurons, while the total number of c-Fos+ cells was decreased (n = 10/group, two-tailed unpaired t test, **p < 0.01). l Reduction in freezing in mice after chemogenetic activation of GABAergic interneurons in the BLA (n = 16/group, two-tailed unpaired t test, **p < 0.01). All data are presented as the mean ± SEM.