Pathological consequences of long-term mitochondrial oxidative stress in the mouse retinal pigment epithelium.
Oxidative stress in the retinal pigment epithelium (RPE) is hypothesized to be a major contributor to the development of age-related macular degeneration (AMD). Mitochondrial manganese superoxide dismutase (MnSOD) is a critical antioxidant protein that scavenges the highly reactive superoxide radical. We speculated that specific reduction of MnSOD in the RPE will increase the level of reactive oxygen species in the retina/RPE/choroid complex leading to pathogenesis similar to geographic atrophy. To test this hypothesis, an Sod2-specific hammerhead ribozyme (Rz), delivered by AAV2/1 and driven by the human VMD2 promoter was injected subretinally into C57BL/6J mice. Dark-adapted full field electroretinogram (ERG) detected a decrease in the response to light. We investigated the age-dependent phenotypic and morphological changes of the outer retina using digital fundus imaging and SD-OCT measurement of ONL thickness. Fundus microscopy revealed pigmentary abnormalities in the retina and these corresponded to sub-retinal and sub-RPE deposits seen in SD-OCT B-scans. Light and electron microscopy documented the localization of apical deposits and thickening of the RPE. In RPE flat-mounts we observed abnormally displaced nuclei and regions of apparent fibrosis in the central retina of the oldest mice. This region was surrounded by enlarged and irregular RPE cells that have been observed in eyes donated by AMD patients and in other mouse models of AMD.
Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL 32610, USA.
Soo-jung
SJ
Seo
Mark P
MP
Krebs
Haoyu
H
Mao
Kyle
K
Jones
Mandy
M
Conners
Alfred S
AS
Lewin
D000818:Animals; D000229:Dependovirus / Q000235:genetics; D004195:Disease Models, Animal; D004596:Electroretinography; D005451:Fluorescein Angiography; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D020868:Gene Silencing / Q000502:physiology; D005822:Genetic Vectors; D057092:Geographic Atrophy / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000201:enzymology*; D018384:Oxidative Stress; D016337:RNA, Catalytic / Q000235:genetics; D055213:Retinal Pigment Epithelium / Q000201:enzymology / Q000648:ultrastructure*; D013482:Superoxide Dismutase / Q000235:genetics*; D041623:Tomography, Optical Coherence; D000099156:Superoxide Dismutase 2
D016337:RNA, Catalytic; D013482:Superoxide Dismutase; D000099156:Superoxide Dismutase 2; C427204:hammerhead ribozyme