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Author Notes:

Correspondence: John M. Nickerson, Department of Ophthalmology, Emory University, B5602, 1365B Clifton Road, NE, Atlanta, GA 30322; litjn@emory.edu.

Disclosure: Y. Jiang, None; X. Qi, None; M.A. Chrenek, None; C. Gardner, None; J.H. Boatright, None; H.E. Grossniklaus, None; J.M. Nickerson, None.


Research Funding:

Supported by Research to Prevent Blindness, Inc., The Katz Foundation, US National Institutes of Health Grants R01EY016470, R01EY014026, R01EY021592, and P30EY006360, and the Neurosciences Initiative of Emory University.


  • RPE
  • image analysis
  • retinal degeneration

Functional Principal Component Analysis Reveals Discriminating Categories of Retinal Pigment Epithelial Morphology in Mice


Journal Title:

Investigative Ophthalmology & Visual Science


Volume 54, Number 12


, Pages 7274-7283

Type of Work:

Article | Final Publisher PDF


Purpose. To determine whether multivariate, functional principal component analysis of the size and shape of retinal pigment epithelial (RPE) cell morphology allows discrimination of mouse RPE genotypes and age. Methods. Flatmounts of RPE sheets obtained from C57BL/6J (n = 50) and rd10 (n = 61) mice at postnatal days 30 to 720 were stained for zonula occludens-1 (ZO-1) and imaged with confocal microscopy. A total of 111 flatmounts were prepared. Twenty-one morphometric measurements were made on tiled, composite images of complete flatmounts, including cell location, area, and eccentricity, using automated image analysis software for quantitatively measuring cell phenotypes. Results. In young (≤61-day-old) C57BL/6J mice, the RPE morphology resembled a regular hexagonal array of cells of uniform size throughout the retina, except near the ciliary body, where the shapes of RPE resembled a soft network. Old (≥180-day-old) C57BL/6J eyes had a subpopulation of large cells. A clear disruption of the regular cell size and shape appeared in rd10 mutants. Aspect ratio and cell area gave rise to principal components that predictively classified mouse age and genotype. Conclusions. Quantitative differences in the RPE sheet morphology allowed discrimination of rd10 from C57BL/6J strains despite the confounding effect of aging. This has implications for RPE sheet morphology as a potential early biomarker for diagnosis of eye disease and prognosis of the eye at early stages when disease is subtle. We conclude that an RPE cell's area and aspect ratio are very early quantitative indicators that predict progression to advanced RPE disease as manifested in rd10.

Copyright information:

© 2013 The Association for Research in Vision and Ophthalmology, Inc.

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