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. 2023 Jan 16;28(3):1090–1100. doi: 10.1038/s41380-022-01922-y

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© The Author(s) 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. 5 — A, B The distribution pattern of the LS GABAergic projection. A Schematics and timeline of antegrade virus (AAV2/9-DIO-EGFP) tracing. B Schematic summary of mapped recipient brain regions of LS projection fibers with the distribution pattern from whole-brain tracing. C–O By using chemogenetic manipulation, LS GABAergic projections to HDB, LH, PAG were indirectly activated in behavioral experiments. C Experimental scheme of virus injections and behavior tests. Schematics of the stereotaxic surgery targeting the (D) LS-HDB, (H) LS-LH, and (L) LS-PAG circuits. Coronal sections illustrating hM4Di mCherry-positive HDB, LH and PAG neurons at the injection site. Scale bar, 100 μm. E–G Behavioral results for LS-HDB activation. E Withdrawal threshold of the hind paw measured with the von Frey test. F OFT (Left) total distance and (Right) percentage of time in center. G EPM (Left) total entries in arms and (Right) time in open arms. I–K Same as (E–G) but for LS-LH activation. M–O Same as (E–G) but for LS-PAG activation. *P < 0.05, **P < 0.01. ns, no significant difference (P > 0.05). Data are presented as the means ± SEM. For further details of statistical data analysis see Table S1. DM, dorsomedial hypothalamus; HDB, horizontal limb of the diagonal band; HPC, hippocampus; LH, lateral hypothalamus; LPO and MPO, lateral and medial preoptic area; NAc, nucleus accumbens; PAG, periaqueductal gray; VDB, vertical limb of the diagonal band; VMH, ventromedial hypothalamus; VTA, ventral tegmental area.