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Author Notes:

Correspondence and requests for materials should be addressed to L.C.S.T. (email: lawrenct@tcd.ie) or W.D.S. (email: william.stamer@duke.edu) or P.H. (email: pete.humphries@tcd.ie)

L.C.S.T. and E.R.-T. contributed equally to this work.

All authors reviewed the manuscript.

L.C.S.T. performed PCR arrays, RNAi knockdown, gene expression analysis in SCEC, IHC, TEER and Papp measurements, and wrote the main manuscript text; E.R.-T. performed mouse intracameral injections, iPerfusion, gene expression analysis in TM cells and contributed to manuscript preparation; J.M.S. contributed to experimental design, performed statistical analysis for iPerfusion and contributed to manuscript preparation; P.C. performed RNAi knockdown, mouse intracameral injection and assisted with iPerfusion; D.C. performed mouse IHC and protein inhibition analysis in mouse; E.L.-D. performed TEM; C.F.-K. performed TEM; K.P. provided SCEC lines; M.M.H. performed gene sequencing and contributed to siRNA design; A.-S.K. assisted with PCR array analysis and contributed to manuscript preparation; J.O’C assisted with iPerfusion and performed cell viability experiments; J.J.C. provided non-human primate anterior segment paraffin sections; A.T.R. contributed to the interpretation of physiological data and statistical analysis, and provided revisions to the manuscript; C.R.E. contributed to the interpretation of physiological data and statistical analysis, and provided revisions to the manuscript; C.O’B. contributed to manuscript preparation; M.L. contributed to manuscript preparation; M.C. contributed to manuscript preparation; W.D.S. conceived the concept and assisted with experimental design, provided cell lines and contributed to manuscript preparation; D.R.O. assisted with experimental design, iPerfusion and contributed to manuscript preparation; P.H. conceived the concept and contributed to experimental design and writing of the manuscript.

We would like to thank C. Woods and C. Murray for animal husbandry. We thank Elke Kretshmar, Hong Nguyen and Britta Bäckermann for their technical assistance with TEM, Jörg Pekarsky for preparing the schematic drawing and Marco Gößwein for preparing the final version of Figure 1.

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Research Funding:

Work at the Ocular Genetics Unit at the University of Dublin, Trinity College, was supported by the European Research Council ERC-2012-AdG.

The Unit also receives support from Science Foundation Ireland.

Work at Duke University was supported by grants from the US National Institutes of Health (EY022359 and EY019696) and at Imperial College London by Fight for Sight UK (Ref 1385), the US National Institutes of Health (EY022359 and EY019696), and the UK Engineering and Physical Sciences Research Council (EP/J010499/1).

Keywords:

  • Science & Technology
  • Multidisciplinary Sciences
  • Science & Technology - Other Topics
  • INNER-WALL ENDOTHELIUM
  • TRABECULAR MESHWORK
  • AQUEOUS-HUMOR
  • MONKEY EYE
  • INTERENDOTHELIAL JUNCTIONS
  • INTRAOCULAR-PRESSURE
  • FREEZE-FRACTURE
  • PORE DENSITY
  • CELLS
  • BARRIER

Enhancement of Outflow Facility in the Murine Eye by Targeting Selected Tight-Junctions of Schlemm's Canal Endothelia

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Journal Title:

Scientific Reports

Volume:

Volume 7

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Type of Work:

Article | Final Publisher PDF

Abstract:

The juxtacanalicular connective tissue of the trabecular meshwork together with inner wall endothelium of Schlemm's canal (SC) provide the bulk of resistance to aqueous outflow from the anterior chamber. Endothelial cells lining SC elaborate tight junctions (TJs), down-regulation of which may widen paracellular spaces between cells, allowing greater fluid outflow. We observed significant increase in paracellular permeability following siRNA-mediated suppression of TJ transcripts, claudin-11, zonula-occludens-1 (ZO-1) and tricellulin in human SC endothelial monolayers. In mice claudin-11 was not detected, but intracameral injection of siRNAs targeting ZO-1 and tricellulin increased outflow facility significantly. Structural qualitative and quantitative analysis of SC inner wall by transmission electron microscopy revealed significantly more open clefts between endothelial cells treated with targeting, as opposed to non-targeting siRNA. These data substantiate the concept that the continuity of SC endothelium is an important determinant of outflow resistance, and suggest that SC endothelial TJs represent a specific target for enhancement of aqueous movement through the conventional outflow system.

Copyright information:

© The Author(s) 2017.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
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