title: Altered expression of genes related to blood-retina barrier disruption in streptozotocin-induced diabetes.
abstract: Disruption of the blood-retina barrier (BRB) is an early phenomenon in preclinical diabetic retinopathy (PCDR). Two vascular permeability pathways may be affected, the paracellular pathway involving endothelial cell tight junctions, and the endothelial transcellular pathway mediated by endocytotic vesicles (caveolae). The relative contribution of both pathways to vascular permeability in PCDR is unknown. We compared transcription levels in entire rat retina of genes related to these pathways between control conditions and after 6 and 12 weeks of streptozotocin-induced diabetes, as well as in bovine retinal endothelial cells (BRECs) exposed to VEGF and bovine retinal pericytes (BRPCs), using real-time quantitative RT-PCR. To confirm endothelial-specificity, immunohistochemical staining was performed in rat retina, and mRNA transcript levels were compared between BRECs and BRPCs. mRNA and protein of most paracellular transport-related genes were specifically expressed by retinal endothelial cells, whereas vesicle transport-related mRNA and proteins were present in various retinal cell types, including endothelial cells. Expression of selected endothelial cell tight junction genes and particularly that of occludin and claudin-5 was reduced in the diabetic retina and in BRECs after exposure to VEGF. Expression of 6 out of 11 vesicular transport-related genes was upregulated after induction of diabetes. Of these, only plasmalemma vesicle-associated protein (PV-1) was exclusively expressed in BRECs and not in BRPCs. PV-1 transcription was markedly induced in diabetic retina and by VEGF in BRECs. Caveolin-1 immunostaining was primarily found in the retinal vasculature, and its mRNA levels in BRECs were highly abundant and VEGF-inducible. Whereas the endothelial tight junction genes occludin and claudin-5 showed a transient downregulation, we observed long-term upregulation in diabetic retina and VEGF-induced expression in BRECs of the vesicular transport-related genes caveolin-1 and PV-1. The altered gene expression profiles observed in this study suggest a transient induction of the paracellular pathway and prolonged involvement of transcellular endothelial transport mechanisms in the increased permeability of retinal capillaries in PCDR.
authors.affiliation: Ocular Angiogenesis Group, Department of Ophthalmology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands. i.klaassen@amc.uva.nl
authors.forename: Ingeborg
authors.initials: I
authors.lastname: Klaassen
authors.forename: John M
authors.initials: JM
authors.lastname: Hughes
authors.forename: Ilse M C
authors.initials: IM
authors.lastname: Vogels
authors.forename: Casper G
authors.initials: CG
authors.lastname: Schalkwijk
authors.forename: Cornelis J F
authors.initials: CJ
authors.lastname: Van Noorden
authors.forename: Reinier O
authors.initials: RO
authors.lastname: Schlingemann
mesh_terms: D022005:Adherens Junctions / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001813:Blood-Retinal Barrier / Q000502:physiology*; D002199:Capillary Permeability / Q000235:genetics; D002417:Cattle; D002478:Cells, Cultured; D030561:Databases, Nucleic Acid; D003921:Diabetes Mellitus, Experimental / Q000235:genetics* / Q000378:metabolism; D003930:Diabetic Retinopathy / Q000235:genetics* / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism; D005136:Eye Proteins / Q000096:biosynthesis / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology; D020286:Pericytes / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000378:metabolism; D019108:Tight Junctions / Q000235:genetics / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D033921:Vesicular Transport Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D005136:Eye Proteins; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D033921:Vesicular Transport Proteins