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. 2019 Feb 26;9:2830. doi: 10.1038/s41598-019-39063-y

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© The Author(s) 2019

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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Distribution of ORX-immunoreactive (IR) neurons projecting to the brainstem arousal areas. (A) Schematic drawings of three sections which represent the approximate posterior levels to bregma −2.54 mm (rostral), −2.92 mm (middle) and −3.36 mm (Caudal). (B) Photomicrographs showing the injection sites of BDA into the LPB (B1-F1) and CTb into the VTA (B2), DR (C2), PPT (D2), LDT (E2) and LC (F2). (B3- F5) Line drawings showing the distribution of CTb-IR neurons (green filled circles), ORX-IR neurons (blue filled circles), and ORX and CT- double-IR neurons (red stars) in the three regions of hypothalamus. The distance (mm) behind the bregma is noted at the bottom. Scale bars, 500 μm. CTb; cholera toxin b subunit; ORX: orexin; IR: immunoreactive; VTA: ventral tegmental area; DR: dorsal raphe; PPT: pedunculopontine nucleus; LDT: laterodorsal tegmental nucleus; LC: locus ceruleus; BDA: biotinylated dextranamine.