Shear stress-triggered nitric oxide release from Schlemm’s canal cells

Nicole E. Ashpole, Darryl R. Overby, C. Ross Ethier, W. Daniel Stamer

Research output: Contribution to journalArticlepeer-review

65 Scopus citations

Abstract

PURPOSE. Endothelial nitric oxide (NO) synthase is regulated by shear stress. At elevated intraocular pressures when the Schlemm’s canal (SC) begins to collapse, shear stress is comparable with that in large arteries. We investigated the relationship between NO production and shear stress in cultured human SC cells.

METHODS. Schlemm’s canal endothelial cells isolated from three normal and two glaucomatous human donors were seeded into Ibidi flow chambers at confluence, cultured for 7 days, and subjected to steady shear stress (0.1 or 10 dynes/cm2) for 6, 24, or 168 hours. Cell alignment with flow direction was monitored, and NO production was measured using 4-amino-5-methylamino-2′,7′-difluorofluorescein (DAF-FM) and Griess reagents. Human trabecular meshwork (TM) and umbilical vein endothelial cells (HUVECs) were used as controls.

RESULTS. Normal SC strains aligned with the direction of flow by 7 days. Comparing 0.1 vs. 10 dynes/cm2, NO levels increased by 82% at 24 hours and 8-fold after 7 days by DAF-FM, and similar results were obtained with Griess reagent. Shear responses by SC cells at 24 hours were comparable with HUVECs, and greater than TM cells, which appeared shear-insensitive. Nitric oxide production by SC cells was detectable as early as 6 hours and was inhibited by 100 lM nitro-L-arginine methyl ester. Two glaucomatous SC cell strains were either unresponsive or lifted from the plate in the face of shear.

CONCLUSIONS. Shear stress triggers NO production in human SC cells, similar to other vascular endothelia. Increased shear stress and NO production during SC collapse at elevated intraocular pressures may in part mediate IOP homeostasis.

Original languageEnglish (US)
Pages (from-to)8067-8076
Number of pages10
JournalInvestigative Ophthalmology and Visual Science
Volume55
Issue number12
DOIs
StatePublished - Oct 24 2014
Externally publishedYes

Keywords

  • Aqueous humor
  • Conventional outflow
  • Glaucoma
  • Intraocular pressure
  • eNOS

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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