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

Publication ,  Journal Article
Ashpole, NE; Overby, DR; Ethier, CR; Stamer, WD
Published in: Invest Ophthalmol Vis Sci
November 13, 2014

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/cm(2)) 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/cm(2), 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 μM 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.

Duke Scholars

Published In

Invest Ophthalmol Vis Sci

DOI

EISSN

1552-5783

Publication Date

November 13, 2014

Volume

55

Issue

12

Start / End Page

8067 / 8076

Location

United States

Related Subject Headings

  • Trabecular Meshwork
  • Stress, Mechanical
  • Shear Strength
  • Ophthalmology & Optometry
  • Nitric Oxide Synthase
  • Nitric Oxide
  • Middle Aged
  • Male
  • Intraocular Pressure
  • Humans
 

Citation

APA
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Ashpole, N. E., Overby, D. R., Ethier, C. R., & Stamer, W. D. (2014). Shear stress-triggered nitric oxide release from Schlemm's canal cells. Invest Ophthalmol Vis Sci, 55(12), 8067–8076. https://doi.org/10.1167/iovs.14-14722
Ashpole, Nicole E., Darryl R. Overby, C Ross Ethier, and W Daniel Stamer. “Shear stress-triggered nitric oxide release from Schlemm's canal cells.Invest Ophthalmol Vis Sci 55, no. 12 (November 13, 2014): 8067–76. https://doi.org/10.1167/iovs.14-14722.
Ashpole NE, Overby DR, Ethier CR, Stamer WD. Shear stress-triggered nitric oxide release from Schlemm's canal cells. Invest Ophthalmol Vis Sci. 2014 Nov 13;55(12):8067–76.
Ashpole, Nicole E., et al. “Shear stress-triggered nitric oxide release from Schlemm's canal cells.Invest Ophthalmol Vis Sci, vol. 55, no. 12, Nov. 2014, pp. 8067–76. Pubmed, doi:10.1167/iovs.14-14722.
Ashpole NE, Overby DR, Ethier CR, Stamer WD. Shear stress-triggered nitric oxide release from Schlemm's canal cells. Invest Ophthalmol Vis Sci. 2014 Nov 13;55(12):8067–8076.

Published In

Invest Ophthalmol Vis Sci

DOI

EISSN

1552-5783

Publication Date

November 13, 2014

Volume

55

Issue

12

Start / End Page

8067 / 8076

Location

United States

Related Subject Headings

  • Trabecular Meshwork
  • Stress, Mechanical
  • Shear Strength
  • Ophthalmology & Optometry
  • Nitric Oxide Synthase
  • Nitric Oxide
  • Middle Aged
  • Male
  • Intraocular Pressure
  • Humans