Abstract:
PURPOSE:Changes in blood vessel barrier properties contribute to retinal edema in diabetic retinopathy (DR). However, limited data are available to describe the routes of transport for fluids and solutes across the inner blood-retinal barrier (iBRB). In this study, a three-pore model was developed to characterize such routes in normal and elevated glucose levels. METHODS:Diffusive and apparent permeabilities to TAMRA (467 Da), dextran (70 kDa), and LDL (2000 kDa), as well as hydraulic conductivity, were measured across bovine retinal endothelial cell (BREC) monolayers after exposure to normal- and high-glucose media for 6 days. The data were used to develop a model of transport dynamics. Claudin 5 and eNOS Western blot analysis were used to measure changes in expression and phosphorylation. Immunolocalization of ZO-1 and VE-cadherin demonstrated organization of the junctional complex. Apoptosis was measured by TUNEL assay. RESULTS:A three-pore model describes the fractional transport of water and molecular tracers across the retinal endothelial barrier. No change in permeability or hydraulic conductivity was observed after exposure to high glucose, whereas VEGF increased permeability in both normal- and high-glucose environments. The transport results were consistent with ZO-1 and VE-cadherin immunocytochemistry and expression of claudin-5, which were all unaltered by high glucose. CONCLUSIONS:The data describe, for the first time, a model for transport of various size solutes and fluids across endothelial cells of the iBRB. Further, the results support the existence of an indirect pathway by which iBRB permeability is increased through the upregulation of retinal VEGF in response to hyperglycemia.
journal_name
Invest Ophthalmol Vis Scijournal_title
Investigative ophthalmology & visual scienceauthors
Lopez-Quintero SV,Ji XY,Antonetti DA,Tarbell JMdoi
10.1167/iovs.10-5971subject
Has Abstractpub_date
2011-02-28 00:00:00pages
1171-80issue
2eissn
0146-0404issn
1552-5783pii
52/2/1171journal_volume
52pub_type
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