Characterization of peroxy-A2E and furan-A2E photooxidation products and detection in human and mouse retinal pigment epithelial cell lipofuscin.
The nondegradable pigments that accumulate in retinal pigment epithelial (RPE) cells as lipofuscin constituents are considered to be responsible for the loss of RPE cells in recessive Stargardt disease, a blindness macular disorder of juvenile onset. This autofluorescent material may also contribute to the etiology of age-related macular degeneration. The best characterized of these fluorophores is A2E, a compound consisting of two retinoid-derived side arms extending from a pyridinium ring. Evidence indicates that photochemical mechanisms initiated by excitation from the blue region of the spectrum may contribute to the adverse effects of A2E accumulation, with the A2E photooxidation products being damaging intermediates. By studying the oxidation products (oxo-A2E) generated using oxidizing agents that add one or two oxygens at a time, together with structural analysis by heteronuclear single quantum correlation-NMR spectroscopy, we demonstrated that the oxygen-containing moieties generated within photooxidized A2E include a 5,8-monofuranoid and a cyclic 5,8-monoperoxide. We have shown that the oxidation sites can be assigned to the shorter arm of A2E, to the longer arm, or to both arms by analyzing changes in the UV-visible spectrum of A2E, and we have observed a preference for oxidation on the shorter arm. By liquid chromatography-mass spectrometry, we have also detected both monofuran-A2E and monoperoxy-A2E in aged human RPE and in eye cups of Abca4/Abcr-/- mice, a model of Stargardt disease. Because the cytotoxicity of endoperoxide moieties is well known, the production of endoperoxide-containing oxo-A2E may account, at least in part, for cellular damage ensuing from A2E photooxidation.
Jang
Young P
YP
Department of Ophthalmology, Columbia University, New York, New York 10032, USA.
Matsuda
Hiroko
H
Itagaki
Yasuhiro
Y
Nakanishi
Koji
K
Sparrow
Janet R
JR
D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D000368:Aged; D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D004847:Epithelial Cells / Q000166:cytology*; D005663:Furans / Q000737:chemistry*; D006801:Humans; D008027:Light; D008062:Lipofuscin / Q000737:chemistry* / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008875:Middle Aged; D008956:Models, Chemical; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism; D010545:Peroxides / Q000737:chemistry; D010857:Pigment Epithelium of Eye / Q000378:metabolism*; D011726:Pyridinium Compounds / Q000737:chemistry*; D012162:Retinal Degeneration / Q000378:metabolism; D012176:Retinoids / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry; D013997:Time Factors; D014466:Ultraviolet Rays
C112040:A2-E (N-retinylidene-N-retinylethanolamine); C056475:ABCA4 protein, human; D018528:ATP-Binding Cassette Transporters; C512841:Abca4 protein, mouse; D005663:Furans; D008062:Lipofuscin; D010545:Peroxides; D011726:Pyridinium Compounds; D012176:Retinoids; D012997:Solvents; D010100:Oxygen