Fig. 5: Regulation of breathing by activation of CeAGABA neurons projecting to brainstem respiratory centers. | Nature Communications

Fig. 5: Regulation of breathing by activation of CeAGABA neurons projecting to brainstem respiratory centers.

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

Fig. 5

a Schematic of the neural tracing strategy by injecting an anterograde virus encoding mWGA to the CeA to identify postsynaptic neurons located in the brainstem. b–f Representative images showing that postsynaptic neurons of CeAGABA neurons were identified in the LPBN, NTS, LC and preBötC. Scale bars, 20 µm. g Summary diagram illustrating primary outputs of CeAGABA neurons in the brainstem. h Quantification of the numbers of mWGA+ neurons in different brainstem regions (n = 4). i Schematic of the optogenetic strategy. j–s Immunohistochemical validation and photostimulation effects on breathing. Typical images showing CeAGABA neuron axons within the LPBN (j), LC (l), NTS (n), preBötC (p) and RTN (r), as well as fiber traces. Scale bars, 200 µm. Illumination (10 Hz) of the axon terminals of CeAGABA neurons projecting to the LPBN (k), LC (m), NTS (o) and preBötC (q) significantly increased RF and MV but not TV using WBP recordings in quiescent mice. n = 11 mice for LPBN, n = 8 mice for both LC and NTS, n = 9 mice for preBötC, n = 5 mice for RTN. RF: p < 0.0001 for LPBN, p = 0.0235 for LC, p = 0.0384 for NTS, p = 0.0059 for preBötC; MV: p = 0.0004 for LPBN, p = 0.0062 for LC, p = 0.0181 for NTS, p = 0.0119 for preBötC. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001 by two-tailed paired t-test (k, m, o, q, s). All data are presented as the mean ± SEM. Source data is provided as a Source Data file. The materials depicted in (a, g, i) are created in BioRender. Xiaoyi, W. (2025) https://BioRender.com/i81e326. Abbreviations: AP area postrema, Bar barrington’s nucleus, cc central canal, DMNV dorsal motor nucleus of vagus, IRt intermediate reticular nucleus, LC locus coeruleus, LPBN lateral parabrachial nucleus, NA ambiguous nucleus, NTS nucleus tractus solitarius, PCRt parvicellular reticular nucleus, preBötC preBötzinger complex, RTN retrotrapezoid nucleus, RVLM rostral ventrolateral medulla, scp superior cerebellar peduncle.

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