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. 2022 Jul 6;12:11401. doi: 10.1038/s41598-022-15390-5

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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RNAscope in situ hybridization indicates that most of the endopiriform nucleus (EPN) cells are VGLUT1 (Slc17a7) positive and ~ 60% of these cells co-express Oxtr, with a small population of GABAergic Oxtr+ cells. Some neurons that express Oxtr mRNA (red) co-express mRNA for the GABAergic markers (black arrows), Gad1 (A, blue) or Gad2 (B, blue). However, most Oxtr expressing cells were negative for these markers (white arrows). Most of the Oxtr expressing neurons in EPN co-express the markers for VGLUT1 (Slc17a7) (C, E, blue, black arrows) with only a few that expressed only Oxtr (C, F, white arrows). Expression of the marker for VGLUT2 (Slc17a6) (D, blue) was low in EPN but some co-expression with Oxtr was detected (black arrows). The percentage of Oxtr expressing EPN neurons that co-express one of the other markers studied here is shown in E, while the percent of EPN neurons that express one of the other markers that co-express Oxtr is shown in F.