Quantifying Retinal Pigment Epithelium Dysmorphia and Loss of Histologic Autofluorescence in Age-Related Macular Degeneration
| Autor: | Anna V Zarubina, Thomas Ach, Thomas A. Swain, J. Alan Gambril, Christine A. Curcio, Jeffrey D. Messinger, Kenneth R. Sloan, Carrie Huisingh |
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| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Pathology medicine.medical_specialty Phalloidin Fundus Oculi retinal pigment epithelium Fundus (eye) Biology autofluorescence Retina Lipofuscin 03 medical and health sciences chemistry.chemical_compound Macular Degeneration 0302 clinical medicine medicine Humans age-related macular degeneration Retinal pigment epithelium Optical Imaging cytoskeleton General Medicine Macular degeneration medicine.disease Actin cytoskeleton Phenotype eye diseases Tissue Donors Autofluorescence 030104 developmental biology medicine.anatomical_structure Voronoi statistics chemistry Microscopy Fluorescence 030221 ophthalmology & optometry sense organs Bruch Membrane morphometry |
| Zdroj: | Investigative Ophthalmology & Visual Science |
| ISSN: | 1552-5783 0146-0404 |
| Popis: | Purpose Lipofuscin and melanolipofuscin organelles in retinal pigment epithelium (RPE) cells are signal sources for clinical fundus autofluorescence (AF). To elucidate the subcellular basis of AF imaging, we identified, characterized, and quantified the frequency of RPE morphology and AF phenotypes in donor eyes with age-related macular degeneration (AMD). Methods In 25 RPE-Bruch's membrane flat mounts from 25 eyes, we analyzed 0.4-μm z-stack epifluorescence images of RPE stained with phalloidin for actin cytoskeleton. Using a custom ImageJ plugin, we classified cells selected in a systematic unbiased fashion in six phenotypes representing increasing degrees of pathology. For each cell, area, AF intensity, and number of Voronoi neighbors were compared with phenotype 1 (uniform AF, polygonal morphology) via generalized estimating equations. We also analyzed each cell's neighborhood. Results In 29,323 cells, compared with phenotype 1, all other phenotypes, in order of increasing pathology, had significantly larger area, reduced AF, and more variable number of neighbors. Neighborhood area and AF showed similar, but subtler, trends. Cells with highly autofluorescent granule aggregates are no more autofluorescent than others and are in fact lower overall in AF. Pre-aggregates were found in phenotype 1. Phenotype 2, which exhibited degranulation despite normal cytoskeleton, was the most numerous nonhealthy phenotype (16.23%). Conclusions Despite aggregation of granules that created hyperAF aggregates within cells, overall AF on a per cell basis decreased with increasing severity of dysmorphia (abnormal shape). Data motivate further development of subcellular resolution in clinical fundus AF imaging and inform an ongoing reexamination of the role of lipofuscin in AMD. |
| Databáze: | OpenAIRE |
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