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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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Filamentous actin in proximal and distal aqueous drainage tract. (a–d) at z = 10 μm (from the external scleral surface), autofluorescence (AF; green) signal voids (dark) revealed intrascleral plexuses (ISP) lined by filamentous actin (F-actin; red). Dashed circle: location of ostium more proximally in Schlemm’s canal (SC). (e–h) At z = 20 μm, F-actin-positive ciliary muscle (CM) fibers oriented perpendicular to ISP attach at a border region posterior to SC analogous to scleral spur. Curved dashed line: outlines SC lumen within a scleral depression (AF signal void). (i–l) At z = 25 μm, more proximal in SC lumen (j; dashed outline in k), F-actin-positive CM fibers were more prominent and crossed into the region of SC (within dashed outline) within the trabecular meshwork (TM)/inner wall of SC (SCIW) interface. (m–p) At z = 30 μm, vertically-oriented F-actin-positive CM fibers intermingle with cellular F-actin in the TM/IWSC interface region. Horizontal line places an imaginary border between CM and TM/IWSC regions (asterisk). The AF signal (j,n) revealed septae and loculation in SC. (q–t) At z = 35 μm, an F-actin-positive mesh-like labeling pattern was seen in the TM. Red: F-actin (a,e,i,m,q). Green: AF from structural extracellular matrix (b,f,j,n,r). Merged optical sections: (c,g,k,o,s) (red-green). Merged orthogonal reconstructions: (d,h,l,p,t) (red-green). Vertical white line in orthogonal image: z-location of optical section. Curved dashed lines: outline a lumen that continued from ISP to collector channel (CC; dashed circle) to CC opening (CCO) to SC.