April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Patterned Expression of LM-511 and Integrin 6 in Human Schlemm’s Canal Endothelia
Author Affiliations & Notes
  • S. S. VanderWyst
    Biomedical Engineering Graduate Inter-Disciplinary Program,
    University of Arizona, Tucson, Arizona
  • A. T. Read
    Ophthalmology & Vision Science, University of Toronto, Toronto, Ontario, Canada
  • D. R. Overby
    Bioengineering, Imperial College London, London, United Kingdom
  • W. D. Stamer
    Ophthalmology & Vision Science,
    University of Arizona, Tucson, Arizona
  • Footnotes
    Commercial Relationships  S.S. VanderWyst, None; A.T. Read, None; D.R. Overby, None; W.D. Stamer, None.
  • Footnotes
    Support  NIH Cardiovascular Biomedical Engineering Training Grant (HL007955), NEI (E717007)
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 153. doi:https://doi.org/
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      S. S. VanderWyst, A. T. Read, D. R. Overby, W. D. Stamer; Patterned Expression of LM-511 and Integrin 6 in Human Schlemm’s Canal Endothelia. Invest. Ophthalmol. Vis. Sci. 2010;51(13):153. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Aqueous humor outflow resistance is generated near the juxtacanalicular connective tissue and the inner wall endothelium of Schlemm’s canal (SC). Due to a basal-to-apical directed pressure drop across these tissues, we hypothesize that SC cells must strongly adhere to their basement membrane through function-specific laminin heterotrimers and their cognate integrins. The purpose of this study was to characterize the laminin and laminin-specific integrins in Schlemm’s canal basement membrane.

Methods: : Frozen sagittal sections through conventional outflow tissues from human donor eyes were probed with antibodies recognizing several monomers of laminin (laminin α5, β1, γ1, α3, β3, γ2) and laminin-specific integrin pairs (integrin α3, α6, β1, β4). En-face preparations of SC’s inner wall from perfusion-fixed eyes were immunostained for laminin and integrin co-localization. All tissue samples were examined with confocal microscopy and compared to controls. Mature SC cell monolayers were prepared for immunoprecipitation and Western blot to examine specific laminin heterotrimers.

Results: : In situ

Conclusions: : Immunofluorescence, en-face imaging and Western blot show SC expression of LM-511 and integrin α6 in situ and in vitro. The patterned expression suggests that inner wall endothelia utilize integrin α6 to adhere to LM-511 in opposition to the force of basal-apical pressure gradient in vivo.

Keywords: extracellular matrix • cell adhesions/cell junctions • immunohistochemistry 
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