Fluorescence characteristics of lipofuscin fluorophores from human retinal pigment epithelium

Yakovleva, M. A.; Radchenko, A. Sh.; Feldman, T. B.; Kostyukov, Alexey A.; Арбуханова, П.М.; Борзенок, С.А.; Кузьмин, В. А.; Оstrovsky, М. А.

doi:10.1039/c9pp00406h

Cited by 24 publications

(23 citation statements)

References 49 publications

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“…Our results on photooxidation of isolated RPE lipofuscin (Fig. 1F) are consistent with the results of a comparison of fluorescence emission spectra recorded in suspension of human RPE cells isolated from 74 year old eyes affected with AMD and 75 year old eyes without any signs of AMD [63,91]. The fluorescence spectra from the AMD-affected RPE cells obtained with excitation with 488 nm light show that the emission ratio at 550 nm to 600 nm is increased when compared with normal RPE of similar age.…”

Section: Decrease In Long-wavelength Fluorescence During Photooxidation Of Lipofuscin and An Increase In Blue/cyan Or Green To Orange Flusupporting

confidence: 90%

“…Similar but less pronounced changes could be seen for emission at 550 nm. We were interested in fluorescence changes at that wavelength because it has been reported that the ratio of fluorescence intensity at 550 nm to 600 nm is increased in RPE cells from eyes affected by age-related macular degeneration (AMD) in comparison with RPE cells isolated from healthy eyes of similar age [63]. The fluorescence was induced by excitation with 488 nm light.…”

Section: Exposure Of Rpe Lipofuscin To Visible Light Results In a Decrease Of Longwavelength Fluorescence And An Increase In The Short-wamentioning

confidence: 99%

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Photodegradation of Lipofuscin in Suspension and in ARPE-19 cells and the Similarity of Fluorescence of the Photodegradation Product with Oxidized Docosahexaenoate

Różanowska¹,

Rozanowski²

2021

Preprint

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Retinal lipofuscin accumulates with age in the retinal pigment epithelium (RPE) where its fluorescence properties are used to assess the retinal health. It was observed that there is a decrease in lipofuscin fluorescence above the age of 75 years and in early stages of age-related macular degeneration (AMD). The purpose of this study was to investigate the response of lipofuscin isolated from human RPE, and lipofuscin-laden-cells to visible light, and determine whether an abundant component of lipofuscin, docosahexaenoate (DHA) can contribute to lipofuscin fluorescence upon oxidation. Exposure of lipofuscin to visible leads to a decrease of its long-wavelength fluorescence at about 610 nm with concomitant growth of the short-wavelength fluorescence. The emission spectrum of photodegraded lipofuscin exhibits similarity with that of oxidized DHA. Exposure to light of lipofuscin-laden cells leads to loss of lipofuscin granules from cells, while retaining cell viability. The spectral changes of fluorescence in lipofuscin-laden cells resemble those seen during photodegradation of isolated lipofuscin. Our results demonstrate that fluorescence emission spectra together with quantitation of intensity of long-wavelength fluorescence can serve as a marker useful for lipofuscin quantification and for monitoring its oxidation, thereby useful for screening the retina for increased oxidative damage and early AMD-related changes.

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Section: Decrease In Long-wavelength Fluorescence During Photooxidation Of Lipofuscin and An Increase In Blue/cyan Or Green To Orange Flusupporting

confidence: 90%

Section: Exposure Of Rpe Lipofuscin To Visible Light Results In a Decrease Of Longwavelength Fluorescence And An Increase In The Short-wamentioning

confidence: 99%

Photodegradation of Lipofuscin in Suspension and in ARPE-19 cells and the Similarity of Fluorescence of the Photodegradation Product with Oxidized Docosahexaenoate

Różanowska¹,

Rozanowski²

2021

Preprint

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“…However, the overwhelming density of brightly autofluorescent debris, such as lipofuscin found near the pial surface, were sometimes detected as false positives by Seg-CNN (Supplementary Figure 3B). We suggest that researchers using this software avoid areas with dense debris or lipofuscin, or at least exclude these regions from analysis, although this can be difficult when imaging in aged tissue (Moreno-García et al, 2018; A. Yakovleva et al, 2020).…”

Section: Resultsmentioning

confidence: 99%

“…We suggest that researchers using this software avoid areas with dense debris or lipofuscin, or at least exclude these regions from analysis, although this can be difficult when imaging in aged tissue (Moreno-García et al, 2018;A. Yakovleva et al, 2020).…”

Section: Cnn-based Analysis Retains Tracking Ability Despite Changes In Image Qualitymentioning

confidence: 99%

Automated in vivo tracking of cortical oligodendrocytes

Call

Sulam

et al. 2021

Preprint

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Oligodendrocytes exert a profound influence on neural circuits by accelerating axon potential conduction, altering excitability and providing metabolic support. As oligodendrogenesis continues in the adult brain and is essential for myelin repair, uncovering the factors that control their dynamics is necessary to understand the consequences of adaptive myelination and develop new strategies to enhance remyelination in diseases such as multiple sclerosis. Unfortunately, few methods exist for analysis of oligodendrocyte dynamics, and even fewer are suitable for in vivo investigation. Here, we describe the development of a fully automated cell tracking pipeline using convolutional neural networks (Oligo-Track) that provides rapid volumetric segmentation and tracking of thousands of cells over weeks in vivo. This system reliably replicated human analysis, outperformed traditional analytic approaches, and extracted injury and repair dynamics at multiple cortical depths, establishing that oligodendrogenesis after cuprizone-mediated demyelination is suppressed in deeper cortical layers. Volumetric data provided by this analysis revealed that oligodendrocyte soma size progressively decreases after their generation, and declines further prior to death, providing a means to predict cell age and eventual cell death from individual time points. This new CNN-based analysis pipeline offers a rapid, robust method to quantitatively analyze oligodendrocyte dynamics in vivo, which will aid in understanding how changes in these myelinating cells influence circuit function and recovery from injury and disease.

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“…Lipofuscin (LF) is a cellular metabolic by-product resulting from incomplete lysosomal degradation of shed photoreceptor outer segments by the RPE and represent the primary substance imaged with short-wavelength autofluorescence (SW-AF). [3] At the start of phototransduction, 11cis-retinal chromophore of rhodopsin absorbs a photon of light and isomerizes to all-transretinaldehyde [Figure 1]. This form may be released from the activated rhodopsin and react with phosphatidylethanolamine (PE), also in the disc membrane, to form N-retinylidene-PE (NRPE).…”

Section: Fundamentals Of Autofluorescence Molecular Originmentioning

confidence: 99%

Fundus Autofluorescence and Clinical Applications

Pole

Ameri

2021

JOVR

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Fundus autofluorescence (FAF) has allowed in vivo mapping of retinal metabolic derangements and structural changes not possible with conventional color imaging. Incident light is absorbed by molecules in the fundus, which are excited and in turn emit photons of specific wavelengths that are captured and processed by a sensor to create a metabolic map of the fundus. Studies on the growing number of FAF platforms has shown each may be suited to certain clinical scenarios. Scanning laser ophthalmoscopes, fundus cameras, and modifications of these each have benefits and drawbacks that must be considered before and after imaging to properly interpret the images. Emerging clinical evidence has demonstrated the usefulness of FAF in diagnosis and management of an increasing number of chorioretinal conditions, such as agerelated macular degeneration, central serous chorioretinopathy, retinal drug toxicities, and inherited retinal degenerations such as retinitis pigmentosa and Stargardt disease. This article reviews commercial imaging platforms, imaging techniques, and clinical applications of FAF.

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Fluorescence characteristics of lipofuscin fluorophores from human retinal pigment epithelium

Abstract: Lipofuscin granules accumulate in the retinal pigment epithelium (RPE) with age, especially in patients with visual diseases, including progressive age-related macular degeneration (AMD).

Cited by 24 publications

References 49 publications

Photodegradation of Lipofuscin in Suspension and in ARPE-19 cells and the Similarity of Fluorescence of the Photodegradation Product with Oxidized Docosahexaenoate

Photodegradation of Lipofuscin in Suspension and in ARPE-19 cells and the Similarity of Fluorescence of the Photodegradation Product with Oxidized Docosahexaenoate

Automated in vivo tracking of cortical oligodendrocytes

Fundus Autofluorescence and Clinical Applications

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