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. 2023 Apr 27;14:2435. doi: 10.1038/s41467-023-38180-7

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a, b Changes in expression of AADC mRNA in rodents exposed to CRS and US. Experimental schematics (a left, b left) and qPCR analysis for rats exposed to CRS (a right; n = 4 independent NS rats and n = 4 independent CRS rats, Two-tailed Mann–Whitney U-tests were used to compare differences between groups) and for mice exposed to US (b right; n = 11 independent NS mice and n = 6 independent US mice). NS, not stressed. c, Schematic, and location of the injection of AAV engineered to overexpress shRNA against AADC. Veh vehicle. d, e In vivo validation of AADC knockdown demonstrated by the expression of EGFP/AADC in the LHb by FISH (d), and the percentage of AADC-positive cells in the LHb by FISH (n = 5 independent sh-Veh mice and n = 6 independent sh-AADC mice) (e). f Experimental paradigm for behavioural assays. g–j Effect of AADC knockdown in the LHb on animals’ body weight (n = 11 independent sh-Veh mice and n = 11 independent sh-AADC mice) (g), immobility time in the TST (n = 18 independent sh-Veh mice and n = 18 independent sh-AADC mice) (h), percentage of sucrose preference (i) and total fluid uptake (j) in the SPT (n = 16 independent sh-Veh mice and n = 16 independent sh-AADC mice). k–n Effect of AADC knockdown in the LHb on VTA dopaminergic and RMTg GABAergic neuronal activity. Expression of c-Fos/TH in the VTA (k) and c-Fos/GAD1 in the RMTg (m) by FISH. Percentage of c-Fos-expressing TH-positive neurons in the VTA (n = 7 independent sh-Veh mice and n = 7 independent sh-AADC mice) (l) and c-Fos-expressing GAD1-positive neurons in the RMTg (n = 7 independent sh-Veh mice and n = 6 independent sh-AADC mice) (n) by FISH. Unless otherwise stated, statistical comparisons were performed using a two-tailed unpaired t test. *p < 0.05, **p < 0.01 and ***p < 0.001. ns, not significant. Data are presented as the mean values ± s.e.m. Source data are provided as a Source Data file.