May 2006
Volume 47, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2006
Analysis of Donor Eyes From a Best Vitelliform Macular Dystrophy Patient, Homozygous for the Bestrophin W93C Mutation
Author Affiliations & Notes
  • B. Bakall
    University of Arizona, Tucson, AZ
    Department of Ophthalmology and Vision Science,
    Department of Genetics and Pathology, Uppsala University, Uppsala, Sweden
  • R.A. Radu
    Jules Stein Eye Institute, University of California, Los Angeles, CA
  • J.B. Stanton
    University of Arizona, Tucson, AZ
    Department of Ophthalmology and Vision Science,
  • J. Burke
    Department of Ophthalmology, Medical College of Wisconsin, The Eye Institute, Milwaukee, WI
  • B.S. MacKay
    University of Arizona, Tucson, AZ
    Department of Ophthalmology and Vision Science,
  • C. Wadelius
    Department of Genetics and Pathology, Uppsala University, Uppsala, Sweden
  • C. Appelqvist
    Eye Clinic, Mora hospital, Mora, Sweden
  • G.H. Travis
    Jules Stein Eye Institute, University of California, Los Angeles, CA
    Department of Biological Chemistry, University of California, School of Medicine, Los Angeles, CA
  • A.D. Marmorstein
    University of Arizona, Tucson, AZ
    Department of Ophthalmology and Vision Science,
    Optical Sciences Center,
  • Footnotes
    Commercial Relationships  B. Bakall, None; R.A. Radu, None; J.B. Stanton, None; J. Burke, None; B.S. MacKay, None; C. Wadelius, None; C. Appelqvist, None; G.H. Travis, None; A.D. Marmorstein, None.
  • Footnotes
    Support  NIH Grants EY13160, EY14898, Research to Prevent Blindness, Phillip Morris USA, Phillip Morris International and Swedish Research Council
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 5817. doi:
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      B. Bakall, R.A. Radu, J.B. Stanton, J. Burke, B.S. MacKay, C. Wadelius, C. Appelqvist, G.H. Travis, A.D. Marmorstein; Analysis of Donor Eyes From a Best Vitelliform Macular Dystrophy Patient, Homozygous for the Bestrophin W93C Mutation . Invest. Ophthalmol. Vis. Sci. 2006;47(13):5817.

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

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Abstract

Purpose: : To determine the biochemical and histopathologic consequences of Best Vitelliform Macular Dystrophy, in eyes from a donor homozygous for the W93C mutation in bestrophin.

Methods: : Eyes were enucleated post mortem. One eye was fixed in 4% Paraformaldehyde, embedded in paraffin, and sectioned. Sections were stained for histological analysis and immunocytochemistry was performed to localize bestrophin. The remaining eye was frozen directly. Retinal pigment epithelium (RPE) was isolated for biochemical analysis. The expression of bestrophin was confirmed by western blot. Intracellular granules were subject to sucrose gradient sedimentation. Granules were counted using a hemocytometer, examined by electron microscopy and analyzed for A2E and its chemical precursors.

Results: : Histologic analysis confirmed complete degeneration and scarring of the macula. Throughout the eye, photoreceptor degeneration was observed and RPE cells appeared hypertrophic with substantial accumulation of intracellular granules. Fluorescence microscopy indicated substantial autofluorescence of granules which were confirmed to contain A2E and its precursors. While A2E levels per granule were lower than in age matched control eyes, the increase in total granules resulted in a 5–7 fold increase in total A2E/eye in the Best donor eye. Analysis of the granules by electron microscopy indicated a complex multilobular structure that differed substantially from granules isolated from age matched control eyes. Immunohistochemistry for bestrophin showed localization exclusive to the RPE.

Conclusions: : These donor eyes provide, for the first time, the histopathology of eyes homozygous for a bestrophin mutation. The study confirms prior findings, where RPE appear increased in size with lipofuscin accumulation. This is the first report indicating that levels of A2E are increased in Best disease.

Keywords: retinal degenerations: hereditary • macula/fovea • retinal pigment epithelium 
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