title: Blood-retinal barrier disruption and ultrastructural changes in the hypoxic retina in adult rats: the beneficial effect of melatonin administration.
abstract: Reactive changes in astrocytes and Müller cells in the retina of adult rats subjected to hypoxia were investigated. Along with this, the integrity of the blood-retinal barrier (BRB) was assessed using fluorescent and electron-dense tracers. In hypoxic rats, mRNA and protein expression of glial fibrillary acidic protein (GFAP) and aquaporin-4 (AQ4) were significantly increased. AQ4 immunoreactive cells were identified as astrocytes and Müller cells by double immunofluorescence labelling. Another alteration in the hypoxic retina was marked reduction in melatonin content compared to controls. In this connection, administration of exogenous melatonin reduced the tissue concentration of vascular endothelial growth factor (VEGF) and nitric oxide (NO); both were elevated in hypoxic rats. A major structural change in the hypoxic retina was swelling of astrocyte and Müller cell processes but this was noticeably attenuated after melatonin administration. Following an intraperitoneal or intravenous injection of rhodamine isothiocyanate (RhIC) or horseradish peroxidase (HRP), leakage of both tracers was observed in the retina in hypoxic rats but not in the controls, indicating that the functional integrity of the BRB is compromised in hypoxia/reoxygenation. It is suggested that enhanced tissue concentration of VEGF and NO production in the hypoxic retina contribute to increased permeability of the retinal blood vessels. The concurrent up-regulation of AQ4, a water-transporting protein, in astrocytes and Müller cells in hypoxia suggests its involvement in oedema formation. Since melatonin effectively reduced the vascular permeability in the retina of hypoxic rats, as evidenced by reduced leakage of RhIC, we suggest that its administration may be of potential benefit in the management of retinal oedema associated with retinal hypoxia.
authors.affiliation: Department of Anatomy, Yong Loo Lin School of Medicine, Blk MD10, 4 Medical Drive, National University of Singapore, Singapore 117597. antkaurc@nus.edu.sg
authors.forename: C
authors.initials: C
authors.lastname: Kaur
authors.forename: V
authors.initials: V
authors.lastname: Sivakumar
authors.forename: Z
authors.initials: Z
authors.lastname: Yong
authors.forename: J
authors.initials: J
authors.lastname: Lu
authors.forename: W S
authors.initials: WS
authors.lastname: Foulds
authors.forename: E A
authors.initials: EA
authors.lastname: Ling
mesh_terms: D000818:Animals; D000975:Antioxidants; D051401:Aquaporin 4 / Q000235:genetics / Q000378:metabolism; D001253:Astrocytes / Q000378:metabolism; D001813:Blood-Retinal Barrier; D002199:Capillary Permeability / Q000187:drug effects; D004353:Drug Evaluation, Preclinical; D016166:Free Radical Scavengers / Q000493:pharmacokinetics / Q000627:therapeutic use; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism; D006735:Horseradish Peroxidase; D000860:Hypoxia / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D008297:Male; D008550:Melatonin / Q000493:pharmacokinetics / Q000627:therapeutic use*; D008854:Microscopy, Electron; D009569:Nitric Oxide / Q000096:biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000378:metabolism / Q000648:ultrastructure*; D012164:Retinal Diseases / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012235:Rhodamines; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D000975:Antioxidants; C496032:Aqp4 protein, rat; D051401:Aquaporin 4; D016166:Free Radical Scavengers; D005904:Glial Fibrillary Acidic Protein; D012333:RNA, Messenger; D012235:Rhodamines; D042461:Vascular Endothelial Growth Factor A; C467498:vascular endothelial growth factor A, rat; D009569:Nitric Oxide; C027755:rhodamine isothiocyanate; D006735:Horseradish Peroxidase; D008550:Melatonin