Fig. 3: Respiratory and behavioral changes following activation of CeAGABA neurons. | Nature Communications

Fig. 3: Respiratory and behavioral changes following activation of CeAGABA neurons.

From: GABAergic neurons in central amygdala contribute to orchestrating anxiety-like behaviors and breathing patterns

Fig. 3

a Schematic diagram of the chemogenetic strategy by delivering a virus containing hM3Dq into the CeA from Vgat-Cre mice. b Representative image of immunohistochemical detection of hM3Dq expression in CeA neurons. Scale bar, 100 μm. c RNAscope-FISH images showing colocalization of hM3Dq (red) and Slc32a1 (green) mRNA in CeAGABA neurons. Scale bars, 20 μm. d Quantitative analysis of the efficiency and specificity of hM3Dq-mCherry. n = 4 mice. e Representative traces showing that bath application of CNO (5 μM) significantly increased firing in a hM3Dq-expressing CeAGABA neuron, as measured by whole-cell patch clamp slice recordings. f Schematic of the experimental procedure. g Distribution of RF (bin size 0.5 Hz) over 40-min assay in saline and CNO injected groups (n = 13 mice per group). p < 0.0001 for 1.5 and 2 Hz. h Representative plethysmography airflow traces of saline and CNO treated groups. i Accumulative duration spent in active and quiescent states during 40-min plethysmography recordings (n = 9 mice per group). Active: p < 0.0001; Quiescent: p < 0.0001. Percent of time (j, n = 9 mice for each group) and accumulative duration (k, n = 9 mice for each group) of each behavior defined in (h). Compared to saline group, moving and grooming episodes increased, and sleeping time decreased. Moving, p = 0.0149; Groom, p < 0.0001; Sleep, p = 0.0001. l Percent of time spent on each behavior in every 5-min interval over 40 min (n = 9 mice for each group). Duration of moving (m), grooming (n), sniff (o), eupnea (p) and sleep (q) in every 5 min. n = 9 per group. m, 11–15 min, p = 0.0004; 21–25 min, p = 0.0077. n, 11–15 min, p = 0.0058; 16–20 min, p < 0.0001. p, 11–15 min, p < 0.0001. q, 16–20 min, p = 0.0001; 21–25 min, p < 0.0001; 26–30 min, p = 0.0245; 31–35 min, p = 0.0064. r CNO injection increased high-frequency respiratory episodes. n = 13 mice per group. 20 min, p = 0.0436; 40 min, p = 0.0021; 50 min, p = 0.0100; 60 min, p = 0.0199. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.001 by two-way ANOVA with Bonferroni’s multiple comparisons tests (g, m–r), two-tailed unpaired t-test (k, groom, eupnea, sleep), two-tailed unpaired t- test with Welch’s correction (i, k, moving, sniff). All data are presented as the mean ± SEM. Source data are provided as a Source Data file. The materials depicted in (a, f) are created in BioRender. Xiaoyi, W. (2025) https://BioRender.com/i81e326.

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