April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Effects of TNF and IL-1β on the Tight Junctions of Human Fetal RPE
Author Affiliations & Notes
  • S. Peng
    Surgery and Ophthal & Visual Science, Yale Univ Sch of Medicine, New Haven, Connecticut
    Ophthalmology, 2nd Hospital of Harbin Medical Unversity, Harbin, China
  • V. Rao
    Surgery and Ophthal & Visual Science, Yale Univ Sch of Medicine, New Haven, Connecticut
  • R. A. Adelman
    Ophthal & Visual Science,
    Yale Univ Sch of Medicine, New Haven, Connecticut
  • L. J. Rizzolo
    Surgery and Ophthal & Visual Science,
    Yale Univ Sch of Medicine, New Haven, Connecticut
  • Footnotes
    Commercial Relationships  S. Peng, None; V. Rao, None; R.A. Adelman, None; L.J. Rizzolo, None.
  • Footnotes
    Support  Yale Endowed Research Fund; Leir Foundation
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 2796. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      S. Peng, V. Rao, R. A. Adelman, L. J. Rizzolo; Effects of TNF and IL-1β on the Tight Junctions of Human Fetal RPE. Invest. Ophthalmol. Vis. Sci. 2010;51(13):2796.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose: : Age-related macular degeneration (AMD) is a chronic disease with a low-grade inflammatory component. Proinflammatory cytokines affect the permeability of tight junctions in a variety of epithelia. This study investigates how tumor necrosis factor (TNF)-α and interleukin (IL)-1β, affect the permeability and selectivity of tight junctions in the retinal pigment epithelium (RPE).

Methods: : Human fetal RPE (hfRPE) was cultured on filters. TNFα (20ng/ml) or IL-1β (20ng/ml) was added to the apical and basal media chambers for 48 hours. Transepithelial electrical resistance (TER) was used to monitor the permeability and selectivity of RPE tight junctions. The relative conductance of Na+ and K+ was estimated from the TER in buffered NaCl or KCL solutions. The claudin composition of RPE tight junctions was examined using real-time RT-PCR to estimate relative mRNA levels, immunoblotting to monitor protein expression, and immunofluorescence to monitor protein distribution.

Results: : TNFα decreased the TER from 675 Ω-cm2 to 102 Ω-cm2. The ratio of the conductance for Na+ /K+ increased from 0.54 to 0.71. By contrast, IL-1β had little effect on TER or relative conductance. The expression of claudin 19 mRNA was more than 20X greater than the other claudin, but was minimally affected by TNFα. TNFα increased the expression of claudin 1, 2, and 3 mRNAs and decreased the expression of claudin 10 mRNA. Other claudins were only slightly affected. Corresponding changes in protein expression were observed by immunoblotting. Although the effects on protein distribution were minimal, TNFα affected structure, giving the tight junctions a jagged appearance in the plane of the monolayer and increasing the number of apical stress fibers. IL-1β had minimal effect on claudin mRNA or protein expression, claudin subcellular localization, or junctional structure.

Conclusions: : Rather than compromise the outer blood-retinal barrier by disrupting RPE tight junctions, TNFα altered the properties of the junctions. The junctions became more permeable to ions and less selective for K+ over Na+. These changes correlated with increased expression of minor claudins and altered tight junction morphology.

Keywords: retinal pigment epithelium • pump/barrier function • inflammation 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×