Retinal vascular changes after glial disruption in rats.
Glial dysfunction is found in a number of retinal vascular diseases but its link with blood-retinal barrier (BRB) breakdown remains poorly understood. The present study tested the hypothesis that glial dysfunction is a major contributor to the BRB breakdown that is a hallmark of retinal vascular diseases. We investigated the specificity of the purportedly selective glial toxin, DL-alpha-aminoadipic acid (DL-alpha-AAA) on different types of ocular cells in vitro and then tested the effect of glial disruption on retinal vasculature after intraocular injection of DL-alpha-AAA or siRNA targeting glutamine synthetase (GS) in rats. DL-alpha-AAA was toxic to astrocytes and Müller cells but not to other types of BRB-related cells in vitro. Subretinal injection of DL-alpha-AAA disrupted retinal glial cells, induced vascular telangiectasis and increased vascular permeability from 4 days to over 2 months post-injection. Vascular changes induced by DL-alpha-AAA were observed predominantly in regions of glial disruption, as reflected by reduced expression of GS and increased expression of glial fibrillary acidic protein and vimentin. Confocal microscopy showed changes in all three layers of the retinal vasculature, which co-localised with areas of Müller cell disruption. Double labeling immunohistochemistry revealed that retinal glial disruption after DL-alpha-AAA injection was accompanied by increased expression of vascular endothelial growth factor and reduced expression of the tight junction protein claudin-5. Intravitreal injection of GS siRNA induced similar changes in Müller cells and BRB breakdown. Our data are consistent with the hypothesis that glial dysfunction is a primary contributor to the BRB breakdown in retinal vascular diseases.
Shen
Weiyong
W
Save Sight Institute, University of Sydney, Sydney, Australia. shenw@med.usyd.edu.au
Li
Shiying
S
Chung
Sook Hyun
SH
Gillies
Mark C
MC
D015074:2-Aminoadipic Acid / Q000633:toxicity; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D001812:Blood-Brain Barrier / Q000473:pathology / Q000503:physiopathology*; D002460:Cell Line; D002478:Cells, Cultured; D062507:Claudin-5; D018691:Excitatory Amino Acid Antagonists / Q000633:toxicity; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D005974:Glutamate-Ammonia Ligase / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009498:Neurotoxins / Q000633:toxicity*; D034622:RNA Interference; D051381:Rats; D012161:Retinal Artery / Q000473:pathology / Q000503:physiopathology*; D031300:Retinal Vasculitis / Q000473:pathology / Q000503:physiopathology*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D014746:Vimentin / Q000378:metabolism
C107466:CLDN5 protein, human; D062507:Claudin-5; D018691:Excitatory Amino Acid Antagonists; D005904:Glial Fibrillary Acidic Protein; D008565:Membrane Proteins; D009498:Neurotoxins; D042461:Vascular Endothelial Growth Factor A; D014746:Vimentin; C467498:vascular endothelial growth factor A, rat; D015074:2-Aminoadipic Acid; D005974:Glutamate-Ammonia Ligase