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. 2023 Feb 24;14:1066. doi: 10.1038/s41467-023-36641-7

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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. 3 — a Schema of rAAV_1/2-pEF1α-DIO-GFP injection in the PAG of OTR-Cre rats. b Graph showing the relation between the number of OT fibers in the vlPAG and the bregma level. Blue area represents SEM. n_fibers = 290, n_rats = 4. c Graph showing the relation between the percentage of OTR neurons in the vlPAG and the bregma level. Blue area represents SEM. n_cells = 528, n_rats = 4. d Correlation between the numbers of OTR neurons and OT fibers within the same slice (n_slice = 64). Each dot represents one analyzed brain section. Pearson r correlation, R² = 0.5745, p < 0.0001 (two-sided), slope = 0.2438. e, f Three-dimensional reconstruction of OTergic contacts with vlPAG OTR neurons. e Overview image showing OTR-neurons (green), OT-fibers (magenta), synaptophysin (SYN, red), and DAPI (blue). f Magnified images showing contacts with or without SYN. White arrowheads indicate co-localization of OT (magenta) and SYN (red), while white arrowheads with an asterisk show a mismatch of OT and SYN. DAPI = blue, OTR = green. g Bar graph showing the percentage of OTR positive (n = 496) and negative (n = 3840) cells receiving OT innervation (<1 µm distance between fibers and cells). n_rats = 4, 8 images per animal, p = 0.0055 (two-sided). h Bar graph showings the percentage of contacts between OT and OTR-positive neurons at somatic and dendritic locations. n_rats = 4, 8 images per animal, p < 0.0001 (two-sided). i Reconstruction of a vGluT2-positive (red) OT fibers (magenta) within the vlPAG. j Bar graph showing that the vast majority of OT fibers within the vlPAG are vGluT2-negative (92.4%). n = 4. k, l 3D reconstruction of contacts between an OTR dendrite and OT fibers. k Co-localization of OT (magenta) and vGluT2 (red) are indicated by white arrowheads. l Mismatch of OT (magenta) and vGluT2 (red) are indicated by white arrowheads with an asterisk. DAPI = blue, OTR = green. n = 4 female rats. Scale bars in order of appearance: 50, 10, 10, 20, and 20 µm. Results are expressed as the mean ± SEM and the individual points of each conditions are represented as white circle. Source data are provided as a Source data file.