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. 2018 Aug 1;8:11540. doi: 10.1038/s41598-018-29915-4

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© The Author(s) 2018

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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lVTA-projecting LDT cholinergic neurons in Runner mice. (A) After two weeks of VWR, 50 nl of RBR was injected into the left side of the lVTA in mice, and RBR-injected mice continued VWR for 2 more weeks. (B) Changes in running distances in RBR-injected mice (n = 6). (C) Photomicrograph showing positive signals for RBR (Red) and nNOS (Blue) in the ipsilateral and contralateral sides of the injection in the LDT. Aq: aqueduct. (D,E) Photomicrographs showing positive signals for nNOS (Blue), RBR (Red) and FosBΔFosB (Green) in the LDT. Arrows and arrowheads indicate lVTA-projecting activated LDT-cholinergic (nNOS⁺ RBR⁺ FosBΔFosB⁺) neurons and lVTA-projecting activated LDT-non-cholinergic (nNOS⁻ RBR⁺ FosBΔFosB⁺) neurons, respectively. Bars: (C) = 50 μm, (D,E) = 20 μm. (F) A table showing the quantitative results of cholinergic, lVTA-projecting activated cholinergic and lVTA-projecting activated non-cholinergic neurons on the ipsilateral and contralateral sides of the injection (n = 6). Quantitative data are presented as the mean ± standard error of the mean (SEM).