title: Effect of Rpe65 knockout on accumulation of lipofuscin fluorophores in the retinal pigment epithelium.
abstract: OBJECTIVE
  In all mammalian species examined to date the retinal pigment epithelium (RPE) has been found to accumulate autofluorescent lysosomal storage bodies (lipofuscin) during senescence. Substantial evidence indicates that retinoids in the RPE-retina complex play a major role in RPE lipofuscin formation. Indeed, at least one RPE lipofuscin fluorophore is derived in part from vitamin A aldehyde. However, the precise mechanisms by which retinoids modulate RPE lipofuscin accumulation have not been elucidated. In mice without a functional Rpe65 gene, isomerization of all-trans- to 11-cis-retinol is blocked. Experiments were performed to determine whether this impairment of retinoid metabolism alters RPE lipofuscin accumulation.
  
  
  METHODS
  RPE lipofuscin fluorophore content was compared in 12- to 13-month-old Rpe65(+/+), Rpe65(+/-), and Rpe65(-/-) mice. Lipofuscin fluorophore content was determined using quantitative fluorometric measurements. RPE lipofuscin content was also estimated with quantitative ultrastructural techniques.
  
  
  RESULTS
  In the Rpe65(-/-) mice, RPE lipofuscin fluorophore accumulation was almost abolished. In addition, a significantly reduced accumulation of lipofuscin fluorophores was also observed in the Rpe65(+/-) animals. The inability of the RPE of)Rpe65(-/-) mice to supply 11-cis-retinal from the RPE to the retinal photoreceptors was accompanied by a massive accumulation of lipid droplets in the RPE that appeared to contain substantial amounts of retinoids.
  
  
  CONCLUSIONS
  These findings indicate that formation of RPE lipofuscin fluorophores is almost completely dependent on a normal visual cycle. The absence of retinal (both all-trans and 11-cis) in Rpe65 knockout mice drastically reduced formation of lipofuscin fluorophores in these animals. Even an excessive accumulation of retinyl fatty acid esters in the RPE of Rpe65 knockout mice did not contribute to lipofuscin accumulation.
authors.affiliation: University of Missouri School of Medicine, Mason Eye Institute, Columbia 65212, USA. katzm@health.missouri.edu
authors.forename: M L
authors.initials: ML
authors.lastname: Katz
authors.forename: T M
authors.initials: TM
authors.lastname: Redmond
mesh_terms: D000818:Animals; D002352:Carrier Proteins; D003592:Cytophotometry; D005136:Eye Proteins / Q000502:physiology*; D005227:Fatty Acids / Q000378:metabolism; D017353:Gene Deletion; D008062:Lipofuscin / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008856:Microscopy, Fluorescence; D010857:Pigment Epithelium of Eye / Q000378:metabolism* / Q000648:ultrastructure; D011506:Proteins / Q000502:physiology*; D012172:Retinaldehyde / Q000378:metabolism; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase
chemical_list: D002352:Carrier Proteins; D005136:Eye Proteins; D005227:Fatty Acids; D008062:Lipofuscin; D011506:Proteins; D012172:Retinaldehyde; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase