April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Retina Proteome Profiling in a Rat Model of Smith-Lemli-Opitz Syndrome Using an Ion-Current-Based Method
Author Affiliations & Notes
  • Chengjian Tu
    Pharmaceut. Sci. & NY State Ctr. of Excellence in Life Sci., SUNY-Buffalo, Buffalo, New York
  • Xiaosheng Jiang
    Pharmaceut. Sci. & NY State Ctr. of Excellence in Life Sci., SUNY-Buffalo, Buffalo, New York
  • Jun Qu
    Pharmaceut. Sci. & NY State Ctr. of Excellence in Life Sci., SUNY-Buffalo, Buffalo, New York
    SUNY Eye Inst., Buffalo, New York
  • Lowell G. Sheflin
    Ophthalmol. & Biochem., SUNY-Buffalo & VAWNYHS (Research Service), Buffalo, New York
  • Joyce E. Young
    Ophthalmol. & Biochem., SUNY-Buffalo & VAWNYHS (Research Service), Buffalo, New York
  • Bruce A. Pfeffer
    Ophthalmol. & Biochem., SUNY-Buffalo & VAWNYHS (Research Service), Buffalo, New York
  • Steven J. Fliesler
    SUNY Eye Inst., Buffalo, New York
    Ophthalmol. & Biochem., SUNY-Buffalo & VAWNYHS (Research Service), Buffalo, New York
  • Footnotes
    Commercial Relationships  Chengjian Tu, None; Xiaosheng Jiang, None; Jun Qu, None; Lowell G. Sheflin, None; Joyce E. Young, None; Bruce A. Pfeffer, None; Steven J. Fliesler, None
  • Footnotes
    Support  NIH RO1 EY007361 (SJF) and R21 DA027528 (JQ); RPB Unrestricted Grant (SJF)
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 3332. doi:
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      Chengjian Tu, Xiaosheng Jiang, Jun Qu, Lowell G. Sheflin, Joyce E. Young, Bruce A. Pfeffer, Steven J. Fliesler; Retina Proteome Profiling in a Rat Model of Smith-Lemli-Opitz Syndrome Using an Ion-Current-Based Method. Invest. Ophthalmol. Vis. Sci. 2011;52(14):3332.

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

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Abstract

Purpose: : Rats treated with AY9944, a selective inhibitor of DHCR7 (3β-hydroxy-Δ7-dehydrocholesterol reductase), undergo progressive retinal degeneration and provide an animal model of Smith-Lemli-Opitz syndrome (SLOS). Conventional proteomic methods suffer from poor proteomic coverage and difficulties in analyzing membrane proteins. Here, we examined proteomic changes in retinas from AY9944-treated and age-matched control rats using a gel-free, comprehensive proteomics approach.

Methods: : Sprague-Dawley rats were treated with AY9944 as previously described (Arch. Ophthalmol. 122:1190, 2004); untreated age-matched rats served as controls. Retinas (N=5 per group) were harvested at ca. 2 mo of age, and subjected to nano-LC/Oribtrap analysis (J. Proteome Res. 8:2838, 2009). Paraffin-embedded eyes from companion animals were subjected to conventional histological and correlative TUNEL analysis.

Results: : >3500 unique proteins were analyzed, with a false-discovery rate (FDR) of 0.1%; the false-positive rate for quantification was ≤2%, indicating accurate profiling. Levels of ≥132 proteins were significantly (p<0.05) altered by AY9944 treatment, representing a diversity of biological processes, e.g., apoptosis, visual transduction, lipid metabolism, vesicular transport, oxidative/stress response, and ion channel activity. ROM1, rhodopsin, and rhodopsin kinase levels were decreased, relative to controls, whereas GFAP levels were increased, consistent with retinal degeneration. Pyknosis and TUNEL-positive staining in treated, but not control, retinas was observed almost exclusively in the outer nuclear layer, indicating photoreceptor apoptosis.

Conclusions: : Multiple proteomic changes in the retina occur in the SLOS rat model during the normal-to-pathological transition period, consistent with previous histological observations as well as transcriptome perturbations in this model (Siddiqui et al.; ARVO 2008, 2009). Thus, beyond the initial insult to cholesterol biosynthesis, proteomic alterations are implicated in the etiology of retinal degeneration in this model, and may have broader relevance to the SLOS disease mechanism.

Keywords: retinal degenerations: cell biology • proteomics • apoptosis/cell death 
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