April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Elevated ocular A2E and bis-retinoid levels in a rat model of Smith-Lemli-Opitz syndrome
Author Affiliations & Notes
  • Steven J Fliesler
    Research Service, VAWNYHS, Buffalo, NY
    Ophthalmology and Biochemistry, University at Buffalo and SUNY Eye Institute, Buffalo, NY
  • Christopher C Goulah
    Research Service, VAWNYHS, Buffalo, NY
    Ophthalmology and Biochemistry, University at Buffalo and SUNY Eye Institute, Buffalo, NY
  • Bruce A Pfeffer
    Research Service, VAWNYHS, Buffalo, NY
    Ophthalmology and Biochemistry, University at Buffalo and SUNY Eye Institute, Buffalo, NY
  • Keiko Ueda
    Ophthalmology and Pathology & Cell Biology, Columbia University CP&S, New York, NY
  • Janet R Sparrow
    Ophthalmology and Pathology & Cell Biology, Columbia University CP&S, New York, NY
  • Footnotes
    Commercial Relationships Steven Fliesler, None; Christopher Goulah, None; Bruce Pfeffer, None; Keiko Ueda, None; Janet Sparrow, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 1707. doi:
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    • Get Citation

      Steven J Fliesler, Christopher C Goulah, Bruce A Pfeffer, Keiko Ueda, Janet R Sparrow; Elevated ocular A2E and bis-retinoid levels in a rat model of Smith-Lemli-Opitz syndrome. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1707.

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

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Abstract

Purpose: We previously showed that the retinal pigment epithelium (RPE) in the AY9944-induced rat model of Smith-Lemli-Opitz syndrome (SLOS) exhibits marked accumulation of phagosomes and other lipid/membrane inclusions, compared to age-matched control rats. We hypothesized that the RPE in this model would contain substantially elevated amounts of A2E and other bis-retinoids, relative to controls, which might contribute to the observed pathology. We tested this hypothesis in the present study.

Methods: Sprague-Dawley rats were treated with AY9944 to generate the SLOS rat model, as previously described (Fliesler et al., Arch Ophthalmol. 2004); age-matched rats without AY9944 treatment served as controls. At ca. 10-11 wks postnatal, rat were euthanized and eyes (N=4 per group/treatment) were harvested: for biochemical analysis, eyes were flash frozen in liquid nitrogen and stored at -80oC, while for histological analysis eyes were formalin-fixed and stored at 4oC. Analysis of A2E and related bis-retinoids as well as all-trans retinal was performed by HPLC as previously described (Sparrow et al., Methods Molec Biol., 2010). Formalin-fixed, OCT-embedded eyes were cryosectioned, and frozen sections were mounted/coverslipped without staining; RPE autofluorescence was assessed by confocal scanning laser fluorescence microscopy, using 488 nm excitation and 500-600 nm emission.

Results: Compared to controls, SLOS rat eyes exhibited the following fold-change increases (p<0.05): A2E, 1.52; isoA2E, 2.20; total A2Es, 1.71; all-trans retinal, 1.57. RPE cells in SLOS rat eyes also contained increased numbers of punctate, hyperfluorescent inclusions, compared to age-matched controls, consistent in size and distribution with phagosomes derived from ingested rod outer segment tips.

Conclusions: RPE cells in the SLOS rat model contain elevated levels of A2E and related bis-retinoids, consistent with the observed increase in their phagosome content, compared to untreated controls. These changes may contribute to the retinal dysfunction and degeneration observed in the AY9944-induced SLOS rat model.

Keywords: 701 retinal pigment epithelium • 705 retinoids/retinoid binding proteins • 636 pathobiology  
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