June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Elucidating molecular mechanisms of blood retina barrier permeability
Author Affiliations & Notes
  • Daniel Chao
    Ophthalmology, Bascom Palmer Eye Inst, Univ of Miami, Coral Gables, FL
  • Enrique Salero
    Ophthalmology, Bascom Palmer Eye Inst, Univ of Miami, Coral Gables, FL
  • Yan Wang
    Ophthalmology, Bascom Palmer Eye Inst, Univ of Miami, Coral Gables, FL
  • Claude-Henry Volmar
    Psychiatry, University of Miami, Miami, FL
  • Jeffrey Goldberg
    Ophthalmology, Bascom Palmer Eye Inst, Univ of Miami, Coral Gables, FL
  • Footnotes
    Commercial Relationships Daniel Chao, None; Enrique Salero, None; Yan Wang, None; Claude-Henry Volmar, None; Jeffrey Goldberg, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 5612. doi:
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      Daniel Chao, Enrique Salero, Yan Wang, Claude-Henry Volmar, Jeffrey Goldberg; Elucidating molecular mechanisms of blood retina barrier permeability. Invest. Ophthalmol. Vis. Sci. 2013;54(15):5612.

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

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Abstract

Purpose: To develop and characterize an in vitro model of the blood retina barrier. Disruption of this blood retina barrier is central to a variety of retinal vascular diseases such as diabetic retinopathy and age related macular degeneration. Although some of the key regulators of vascular permeability such as VEGF have been uncovered, the regulation of the retinal vascular permeability remains poorly understood. We have adapted an in vitro assay of blood retina barrier permeability in order to utilize an unbiased high throughput chemical genetic approach to identify novel regulators of blood retina barrier.

Methods: RBE4 cells, a well studied rat brain endothelial cell line are plated in transwell plates in a 96 well format using a standard two chamber assay, and permeability to 70kDa dextrans as well as transendothelial resistance are used as readouts for blood retina barrier permeability. In vivo permeability measurements were also assessed using intravenous injection of 70 kda conjugated dextrans into Sprague Dawley rats after intravitreal injection of candidate molecules.

Results: Addition of VEGF, TNF-alpha, and IL-1 beta, molecules which have been shown to be implicated in blood retina barrier breakdown in ocular disease, increased the permeability of RBE4 monolayers by increasing both permeability to 70 kDa dextran as well as decreasing transendothelial resistance in vitro. Preliminary results of a high throughput screen will be discussed as well as methods to validate these candidates in vivo.

Conclusions: We have developed and validated an in vitro model of the blood retina barrier which is amenable to high throughput screening. This approach will be used in order to identify novel regulators of retinal vascular permeability.

Keywords: 700 retinal neovascularization • 436 blood supply • 499 diabetic retinopathy  
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