Erin Flynn1, Keiko Ueda1, Emily Auran1, Jack M Sullivan2, Janet R Sparrow3. 1. Department of Ophthalmology, Columbia University, New York, New York, United States. 2. Research Service, Veterans Administration Western New York Healthcare System, Buffalo, New York, United States. 3. Department of Ophthalmology, Columbia University, New York, New York, United States Department of Pathology and Cell Biology, Columbia University, New York, New York, United States.
Abstract
PURPOSE: This study was conducted to study correlations among fundus autofluorescence (AF), RPE lipofuscin accumulation, and photoreceptor cell degeneration and to investigate the structural basis of fundus AF spots. METHODS: Fundus AF images (55° lens; 488-nm excitation) and spectral-domain optical coherence tomography (SD-OCT) scans were acquired in pigmented Rdh8(-/-)/Abca4(-/-) mice (ages 1-9 months) with a confocal scanning laser ophthalmoscope (cSLO). For quantitative fundus AF (qAF), gray levels (GLs) were calibrated to an internal fluorescence reference. Retinal bisretinoids were measured by quantitative HPLC. Histometric analysis of outer nuclear layer (ONL) thicknesses was performed, and cryostat sections of retina were examined by fluorescence microscopy. RESULTS: Quantified A2E and qAF intensities increased until age 4 months in the Rdh8(-/-)/Abca4(-/-) mice. The A2E levels declined after 4 months of age, but qAF intensity values continued to rise. The decline in A2E levels in the Rdh8(-/-)/Abca4(-/-) mice paralleled reduced photoreceptor cell viability as reflected in ONL thinning. Hyperautofluorescent puncta in fundus AF images corresponded to photoreceptor cell rosettes in SD-OCT images and histological sections stained with hematoxylin and eosin. The inner segment/outer segment-containing core of the rosette emitted an autofluorescence detected by fluorescence microscopy. CONCLUSIONS: When neural retina is disordered, AF from photoreceptor cells can contribute to noninvasive fundus AF images. Hyperautofluorescent puncta in fundus AF images are attributable, in at least some cases, to photoreceptor cell rosettes. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.
PURPOSE: This study was conducted to study correlations among fundus autofluorescence (AF), RPE lipofuscin accumulation, and photoreceptor cell degeneration and to investigate the structural basis of fundus AF spots. METHODS:Fundus AF images (55° lens; 488-nm excitation) and spectral-domain optical coherence tomography (SD-OCT) scans were acquired in pigmented Rdh8(-/-)/Abca4(-/-) mice (ages 1-9 months) with a confocal scanning laser ophthalmoscope (cSLO). For quantitative fundus AF (qAF), gray levels (GLs) were calibrated to an internal fluorescence reference. Retinal bisretinoids were measured by quantitative HPLC. Histometric analysis of outer nuclear layer (ONL) thicknesses was performed, and cryostat sections of retina were examined by fluorescence microscopy. RESULTS: Quantified A2E and qAF intensities increased until age 4 months in the Rdh8(-/-)/Abca4(-/-) mice. The A2E levels declined after 4 months of age, but qAF intensity values continued to rise. The decline in A2E levels in the Rdh8(-/-)/Abca4(-/-) mice paralleled reduced photoreceptor cell viability as reflected in ONL thinning. Hyperautofluorescent puncta in fundus AF images corresponded to photoreceptor cell rosettes in SD-OCT images and histological sections stained with hematoxylin and eosin. The inner segment/outer segment-containing core of the rosette emitted an autofluorescence detected by fluorescence microscopy. CONCLUSIONS: When neural retina is disordered, AF from photoreceptor cells can contribute to noninvasive fundus AF images. Hyperautofluorescent puncta in fundus AF images are attributable, in at least some cases, to photoreceptor cell rosettes. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.
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