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. 2017 Dec 6;7:17071. doi: 10.1038/s41598-017-16897-y

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© The Author(s) 2017

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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Cellular arrangement in distal aqueous drainage tract walls. 2-photon imaging ex vivo after labeling with: Hoechst 33342 for nuclei (a; C57BL/6 mouse eyes); phalloidin-Alexa-568 for F-actin (a); and anti-alpha-smooth muscle actin antibodies (ASMA in Prox1-GFP mouse eyes; (b–d). (a) A thin cellular layer with flat nuclei lay next to the lumen (dark signal void) of an intrascleral plexus (ISP) connecting with a collector channel. This Prox1-GFP-positive layer (b, low magnification; (c), orthogonal reconstruction; (d), high magnification) was consistent with distal tract endothelium. A multilayered cellular layer external and adjacent to the endothelium had plump nuclei and expressed ASMA but not Prox1-GFP. This ASMA-positive layer was consistent with a contractile smooth muscle layer. It correlated with the ASMA-, calponin- and caldesmon-positive regions that prominently co-localized with F-actin labeling shown in Fig. 9. Double-sided arrows represent ISP lumen diameter measurements nearest the collector channel ostium (CCO).