title: Differential sensitivity of cones to iron-mediated oxidative damage.
abstract: OBJECTIVE
  In this study, the hypothesis that increased intraocular levels of iron cause oxidative damage to the retina was tested.
  
  
  METHODS
  Adult C57BL/6 mice were given an intravitreous injection of saline or 0.10, 0.25, or 0.50 mM FeSO(4). Scotopic electroretinograms (ERGs) were performed 3, 7, and 14 days after injection, and photopic ERGs were performed on day 14. Hydroethidine was used to identify superoxide radicals and lipid peroxidation was visualized by staining for hydroxynonenal (HNE). Retinal cell death was evaluated by TUNEL and measurement of inner nuclear layer (INL) and outer nuclear layer (ONL) thickness. Levels of rhodopsin and cone-opsin mRNA were measured by quantitative real time RT-PCR. Cone density was assessed by peanut agglutinin staining and confocal microscopy.
  
  
  RESULTS
  Compared with retinas in saline-injected eyes, retinas from eyes injected with FeSO(4) showed greater fluorescence after intravenous injection of hydroethidine due to superoxide radicals in photoreceptors, greater photoreceptor staining for HNE, a marker of lipid peroxidation, and increased expression of Heme oxygenase 1, an indicator of oxidative stress. ERG b-wave amplitudes were reduced (photopic > scotopic) in FeSO(4)-injected eyes compared with those in saline-injected eyes. Numerous TUNEL-stained nuclei were seen along the outer border of the ONL, the location of cone cell nuclei, at 1 and 2 days after injection of FeSO(4). In FeSO(4)-injected eyes, the thickness of the ONL, but not the INL, was significantly reduced, and 17 days after injection, there were 3.8- and 2.6-fold reductions in the mRNAs for M-cone and S-cone opsin, respectively, whereas there was no significant difference in rhodopsin mRNA. Confocal microscopy of peanut agglutinin-stained sections showed dose-dependent FeSO(4)-induced cone drop out.
  
  
  CONCLUSIONS
  Increased intraocular levels of FeSO(4) cause oxidative damage to photoreceptors with greater damage to cones than rods. This finding suggests that the oxidative defense system of cones differs from that of rods and other retinal cells, and that cones are more susceptible to damage from the type of oxidative stress imposed by iron.
authors.affiliation: Departments of Ophthalmology and Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21287, USA.
authors.forename: Brian S
authors.initials: BS
authors.lastname: Rogers
authors.forename: Robert C A
authors.initials: RC
authors.lastname: Symons
authors.forename: Keiichi
authors.initials: K
authors.lastname: Komeima
authors.forename: Jikui
authors.initials: J
authors.lastname: Shen
authors.forename: Weihong
authors.initials: W
authors.lastname: Xiao
authors.forename: Mara E
authors.initials: ME
authors.lastname: Swaim
authors.forename: Yuan Yuan
authors.initials: YY
authors.lastname: Gong
authors.forename: Shu
authors.initials: S
authors.lastname: Kachi
authors.forename: Peter A
authors.initials: PA
authors.lastname: Campochiaro
mesh_terms: D000818:Animals; D017209:Apoptosis; D004305:Dose-Response Relationship, Drug; D004596:Electroretinography / Q000187:drug effects; D005296:Ferrous Compounds / Q000633:toxicity*; D015870:Gene Expression / Q000187:drug effects; D051547:Heme Oxygenase-1 / Q000378:metabolism; D020287:In Situ Nick-End Labeling; D007267:Injections; D015227:Lipid Peroxidation / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D018384:Oxidative Stress / Q000187:drug effects*; D012333:RNA, Messenger / Q000378:metabolism; D017949:Retinal Cone Photoreceptor Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D012162:Retinal Degeneration / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012243:Rhodopsin / Q000235:genetics; D017299:Rod Opsins / Q000235:genetics; D013481:Superoxides / Q000378:metabolism; D014822:Vitreous Body
chemical_list: D005296:Ferrous Compounds; D012333:RNA, Messenger; D017299:Rod Opsins; D013481:Superoxides; C020748:ferrous sulfate; D012243:Rhodopsin; D051547:Heme Oxygenase-1