June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Quantified autofluorescence maps of human retinal pigment epithelium in age-related macular degeneration (AMD)
Author Affiliations & Notes
  • Thomas Ach
    Dept of Ophthalmology, Univ of Alabama at Birmingham, Birmingham, AL
  • Anna V Zarubina
    Dept of Ophthalmology, Univ of Alabama at Birmingham, Birmingham, AL
  • Kristen M Hammack
    Computer and Information Sciences, University of Alabama at Birmingham, Birmingham, AL
  • Jeffrey D Messinger
    Dept of Ophthalmology, Univ of Alabama at Birmingham, Birmingham, AL
  • Theodore Smith
    Department of Ophthalmology, NYU Langone Medical Center, New York, NY
  • Kenneth R Sloan
    Dept of Ophthalmology, Univ of Alabama at Birmingham, Birmingham, AL
    Computer and Information Sciences, University of Alabama at Birmingham, Birmingham, AL
  • Christine A Curcio
    Dept of Ophthalmology, Univ of Alabama at Birmingham, Birmingham, AL
  • Footnotes
    Commercial Relationships Thomas Ach, None; Anna Zarubina, None; Kristen Hammack, None; Jeffrey Messinger, None; Theodore Smith, None; Kenneth Sloan, None; Christine Curcio, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 2370. doi:
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    • Get Citation

      Thomas Ach, Anna V Zarubina, Kristen M Hammack, Jeffrey D Messinger, Theodore Smith, Kenneth R Sloan, Christine A Curcio; Quantified autofluorescence maps of human retinal pigment epithelium in age-related macular degeneration (AMD). Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):2370.

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

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Abstract
 
Purpose
 

To map quantified autofluorescence (AF) of human retinal pigment epithelium (RPE) at different stages of AMD relative to age-matched controls with healthy maculas.

 
Methods
 

Human chorioretinal tissue (25 donors (69-95 years) were fixed in paraformaldehyde <4 hr after death. Internal fundus examination revealed different AMD stages: incipient and early (n=18), late atrophic (2), and neovascular AMD (5). Incipient and early AMD had qualitatively similar RPE pathology (Ach ARVO2014). RPE flat-mounts preserved the foveal position. RPE cytoskeleton (labeled with AlexaFluor647-phalloidin) and AF (exc. 460-490 nm, em. > 505 nm) were imaged on a spinning disk fluorescence microscope. AF was normalized to a fluorescence reference (Delori, PMID 22016060). Custom FIJI plugins compared normalized AF to 9 age-matched healthy RPE flatmounts (Ach, PMID 25034602). Difference maps plotted mean log pairwise ratios vs. all controls for each eye (i.e., difference of logs).

 
Results
 

Normalized AF is variable among AMD donor eyes, as in age-matched normal eyes. AF levels in AMD tissues are below levels measured for controls (Figure for example). Only a few eyes show similar or higher AF levels, presumably independent from AMD stage. In 9 of 20 eyes with incipient, early, or late atrophic AMD, AF was particularly lower on the inner slope of the rod ring (< 4 mm eccentricity from the fovea, Curcio PMID 2324310). In remaining eyes no specific sub-regional predilection was discerned. AF was independent of AMD stage.

 
Conclusions
 

This is the first systematic analysis of quantified histological AF in AMD eyes. Normal aging leads to increased AF in the perifovea and near periphery (1.25-2.75 and 2.75-4.25 mm eccentricity per Polyak; Ach, PMID 25034602) reflecting high density of rods. In AMD eyes the perifovea is characterized by noticeably reduced AF, indicating the spatial non-correspondence between areas of high lipofuscin-attributable AF and RPE loss. Unchanged or high AF in some tissues suggests additional factors regulating AF that may be best characterized in longitudinal imaging of genetically defined patient populations.  

 
Representative AF map of a donor with incipient AMD (right) shows reduced AF at the perifoveal region. AMD tissues showed reduced AF throughout the flatmount, as compared to control eyes (left).
 
Representative AF map of a donor with incipient AMD (right) shows reduced AF at the perifoveal region. AMD tissues showed reduced AF throughout the flatmount, as compared to control eyes (left).

 
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