Toward an understanding of bisretinoid autofluorescence bleaching and recovery.

Investigative Ophthalmology & Visual Science
Kazunori YamamotoJanet R Sparrow

Abstract

To understand molecular mechanisms underlying photobleaching of the RPE fluorophores responsible for fundus autofluorescence. ARPE-19 cells were allowed to accumulate the bisretinoid, A2E, and were irradiated at 430 nm. For some experiments, the cells were pretreated with vitamin E or sulforaphane and N-acetylcysteine; samples included A2E-free cells. The cells were analyzed by fluorescence microscopy and ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) analysis. A2E free cells were also irradiated and analyzed. Cell death was quantified by double labeling with a membrane impermeable dye and 4',6'-diamino-2-phenylindole (DAPI). A2E that had accumulated in ARPE-19 cells exhibited irradiation-associated autofluorescence bleaching despite the absence of appreciable cell death. Chromatographic analysis with absorbance, fluorescence, and mass spectrometry detection revealed that irradiation of A2E was associated with A2E photoisomerization, photooxidation, and photodegradation. Pretreatment with vitamin E favored fluorescence recovery; this finding was consistent with a process involving photooxidation. A2E that was not cell-associated underwent irradiation-induced bleaching, but fluorescence recovery was not obse...Continue Reading

Citations

Sep 15, 2015·PloS One·Zhao LiuJanet R Sparrow
Nov 17, 2011·Progress in Retinal and Eye Research·Jennifer J HunterDavid R Williams
Mar 21, 2019·Biomedical Optics Express·Matthias KlemmJens Haueisen
Jan 1, 2014·Journal of Clinical Medicine·Janet R Sparrow, Tobias Duncker
May 7, 2020·Journal of Lipid Research·Hye Jin Kim, Janet R Sparrow
Feb 10, 2021·Journal of Lipid Research·Kim Hye Jin, Janet R Sparrow

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