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. 2021 Sep 24;12:5643. doi: 10.1038/s41467-021-25793-z

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© The Author(s) 2021

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 The white rectangle shows the orientation of the horizontal serial scans, starting from the base of the brain. b Traced blood vessels connect the SCN and OVLT. (N = 6 mice). bi Vasculature reconstruction from traces of blood vessels connecting SCN and OVLT. bii Merged AVP, collagen, SMA, and traced blood vessels. Capillaries emerge independently from the SCNs left and right side (open arrowheads). These form anastomoses in the SCNr (solid arrowhead). Branches exiting from SCNr (solid arrow) course along the 3VF before joining the OVLT capillary bed. biii Explanatory drawing. c Serial optic slices (50 µm) of vasculature between the SCN and OVLT demonstrating continuity of portal capillaries (N = 6 mice). The plates show triplets of images as follows: Left panel = merged AVP (white), collagen (green) and SMA (yellow); red arrows and arrowheads are place markers indicating landmarks for orientation in adjacent slices. Middle panel = blood vessel traces are superimposed on immunochemical results of the left panel; Right panel = drawing identifying structures in the middle panel. Details of the serial plates are as follows: ci–v The left and right SCN capillary beds form anastomoses via capillaries traveling in the 3VF. cvi–viii The traced blood vessels in the SCNr form anastomoses. cix–xi The portal blood vessels (red traces, middle panel) lie at the midline. cx Veins that are not part of the portal system (middle column, white arrowheads) and arteries (white arrows) lie more laterally. cxi–xii The midline blood vessels enter OVLT from its ventral aspect. Abbreviations and colour codes: 3VF floor of the third ventricle, SCNc SCN core, SCNr SCN rostrum, SCNs SCN shell, remaining abbreviations as in Fig. 1. OVLT orange, SCNs and SCNr light blue, SCNc dark blue, 3V magenta, and OC green. Scale bar = 100 µm.