May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Elevated A2E Level in the Sod1-Deficient Mouse, an AMD Animal Model
Author Affiliations & Notes
  • S. Azarian
    Genomics Inst of Novartis Rsrch Fndation, San Diego, California
    Genetics,
  • S. Bohan
    Genomics Inst of Novartis Rsrch Fndation, San Diego, California
    Genetics,
  • T. Hollenbeck
    Genomics Inst of Novartis Rsrch Fndation, San Diego, California
    Analytical Chemistry,
  • X. Zhang
    Genomics Inst of Novartis Rsrch Fndation, San Diego, California
    Genetics,
  • J. Isbell
    Genomics Inst of Novartis Rsrch Fndation, San Diego, California
    Analytical Chemistry,
  • S. Panda
    Regulatory Biology Laboratory, Salk Institute for Biological Studies, La Jolla, California
  • R. Glynne
    Genomics Inst of Novartis Rsrch Fndation, San Diego, California
    Genetics,
  • Footnotes
    Commercial Relationships  S. Azarian, GNF, F; GNF, E; S. Bohan, GNF, F; GNF, E; T. Hollenbeck, GNF, F; GNF, E; X. Zhang, GNF, F; GNF, E; J. Isbell, GNF, F; GNF, E; S. Panda, GNF, C; R. Glynne, GNF, F; GNF, E.
  • Footnotes
    Support  None.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 3427. doi:https://doi.org/
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    • Get Citation

      S. Azarian, S. Bohan, T. Hollenbeck, X. Zhang, J. Isbell, S. Panda, R. Glynne; Elevated A2E Level in the Sod1-Deficient Mouse, an AMD Animal Model. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3427. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Cumulative oxidative stress is thought to be a key pathogenic mechanism in age-related macular degeneration (AMD). Accordingly, mice deficient in Cu-, Zn- superoxide dismutase (Sod1) develop an AMD-like phenotype beyond the age of 10 months (PNAS 103:11282). One source of oxidative stress in AMD is lipofuscin. A2E is a well-known photo-oxidative toxin in lipofuscin. To explore the role of A2E in oxidative AMD pathogenesis, Sod1-/- mice were characterized for ocular A2E content and compared with abcr-/- mice.

Methods: : A 96-well format, LC/MS/MS-based A2E assay was developed. A2E was synthesized and purified by flash chromatography and reverse-phase HPLC. The assay was calibrated with purified A2E spiked into naïve eye extracts. The assay was linear from 1 to 300 pmol of A2E/eye. Purified A2-hexanolamine, generated by the same methodology as for A2E, was used as an internal standard. A2E was extracted from enucleated eyeballs with organic solvent and quantified in pmol/eye +/- SEM. At least 3 animals were used per data point.

Results: : Among 7-month-old animals, A2E content was significantly higher (p = 0.04) in Sod1-/- (20.1 +/- 1.3 pmol/eye) compared with that in Sod1+/+ (13.2 +/- 0.4 pmol/eye); Sod1+/- were not significantly different from Sod1+/+. Five-month-old abcr-/- mice, although younger, had several fold higher A2E level (73.5 +/- 2.6 pmol/eye). A2E level in abcr-/- rose almost linearly between 1 and 8 months from 2.7 to102 pmol/eye; there was relatively little increase by 16 months (116.6 pmol/eye).

Conclusions: : Elevated A2E level is associated with and precedes the AMD phenotype in Sod1-/-. Although this A2E level was substantially lower than that in abcr-/-, it may have become pathogenic in Sod1-/- due to anti-oxidant deficiency. The A2E plateau in senescent abcr-/-, which presumably have a functioning Sod1 enzyme, may explain why there is no gross retinal degeneration in these animals. We are currently generating Sod1:abcr double knockouts to develop an animal model with an earlier AMD phenotype and to explore the role of A2E in AMD pathogenesis.

Keywords: age-related macular degeneration • retinal degenerations: hereditary • ipofuscin 
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