title: Mitochondrial DNA damage and repair in RPE associated with aging and age-related macular degeneration.
abstract: OBJECTIVE
  Mitochondrial DNA (mtDNA) damage may be associated with age-related diseases, such as age-related macular degeneration (AMD). The present study was designed to test whether the frequency of mtDNA damage, heteroplasmic mtDNA mutations, and repair capacity correlate with progression of AMD.
  
  
  METHODS
  Macular and peripheral RPE cells were isolated and cultured from human donor eyes with and without AMD. The stages of AMD were graded according to the Minnesota Grading System. Confluent primary RPE cells were used to test the frequency of endogenous mtDNA damage by quantitative PCR. Mutation detection kits were used to detect heteroplasmic mtDNA mutation. To test the mtDNA repair capacity, cultured RPE cells were allowed to recover for 3 and 6 hours after exposure to H(2)O(2), and repair was assessed by quantitative PCR. The levels of human OGG1 protein, which is associated with mtDNA repair, were analyzed by Western blot.
  
  
  RESULTS
  This study showed that mtDNA damage increased with aging and that more lesions occurred in RPE cells from the macular region than the periphery. Furthermore, mtDNA repair capacity decreased with aging, with less mtDNA repair capacity in the macular region compared with the periphery in samples from aged subjects. Most interestingly, the mtDNA damage was positively correlated with the grading level of AMD, whereas repair capacity was negatively correlated. In addition, more mitochondrial heteroplasmic mutations were detected in eyes with AMD.
  
  
  CONCLUSIONS
  These data show macula-specific increases in mtDNA damage, heteroplasmic mutations, and diminished repair that are associated with aging and AMD severity.
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Texas Medical Branch, Galveston, Texas 77555-1106, USA.
authors.forename: Haijiang
authors.initials: H
authors.lastname: Lin
authors.forename: Haifeng
authors.initials: H
authors.lastname: Xu
authors.forename: Fong-Qi
authors.initials: FQ
authors.lastname: Liang
authors.forename: Hao
authors.initials: H
authors.lastname: Liang
authors.forename: Praveena
authors.initials: P
authors.lastname: Gupta
authors.forename: Anna N
authors.initials: AN
authors.lastname: Havey
authors.forename: Michael E
authors.initials: ME
authors.lastname: Boulton
authors.forename: Bernard F
authors.initials: BF
authors.lastname: Godley
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000502:physiology*; D015153:Blotting, Western; D002478:Cells, Cultured; D004249:DNA Damage; D045647:DNA Glycosylases / Q000378:metabolism; D004252:DNA Mutational Analysis; D004260:DNA Repair / Q000502:physiology*; D004272:DNA, Mitochondrial / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D008268:Macular Degeneration / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D016133:Polymerase Chain Reaction; D055213:Retinal Pigment Epithelium / Q000187:drug effects / Q000378:metabolism*; D014019:Tissue Donors; D055815:Young Adult
chemical_list: D004272:DNA, Mitochondrial; D006861:Hydrogen Peroxide; D045647:DNA Glycosylases; C479058:oxoguanine glycosylase 1, human