Back to Search
Start Over
An In-Vitro Cell Model of Intracellular Protein Aggregation Provides Insights into RPE Stress Associated with Retinopathy
- Source :
- International Journal of Molecular Sciences, Vol 21, Iss 6647, p 6647 (2020), International Journal of Molecular Sciences, Volume 21, Issue 18
- Publication Year :
- 2020
- Publisher :
- MDPI AG, 2020.
-
Abstract
- Impaired cargo trafficking and the aggregation of intracellular macromolecules are key features of neurodegeneration, and a hallmark of aged as well as diseased retinal pigment epithelial (RPE) cells in the eye. Here, photoreceptor outer segments (POS), which are internalized daily by RPE cells, were modified by UV-irradiation to create oxidatively modified POS (OxPOS). Oxidative modification was quantified by a protein carbonyl content assay. Human ARPE-19 cells were synchronously pulsed with POS or OxPOS to study whether oxidatively modified cargos can recapitulate features of RPE pathology associated with blinding diseases. Confocal immunofluorescence microscopy analysis showed that OxPOS was trafficked to LAMP1, LAMP2 lysosomes and to LC3b autophagy vacuoles. Whilst POS were eventually degraded, OxPOS cargos were sequestered in late compartments. Co-localization of OxPOS was also associated with swollen autolysosomes. Ultrastructural analysis revealed the presence of electron-dense OxPOS aggregates in RPE cells, which appeared to be largely resistant to degradation. Measurement of cellular autofluorescence, using parameters used to assess fundus autofluorescence (FAF) in age-related macular disease (AMD) patients, revealed that OxPOS contributed significantly to a key feature of aged and diseased RPE. This in vitro cell model therefore represents a versatile tool to study disease pathways linked with RPE damage and sight-loss.
- Subjects :
- 0301 basic medicine
retina
Retinal Pigment Epithelium
Vacuole
AMD
autofluorescence
lcsh:Chemistry
Macular Degeneration
chemistry.chemical_compound
0302 clinical medicine
lcsh:QH301-705.5
Cells, Cultured
Spectroscopy
LAMP2
LAMP1
Chemistry
Neurodegeneration
General Medicine
Computer Science Applications
Cell biology
medicine.anatomical_structure
RPE
Oxidation-Reduction
Intracellular
proteolysis
autophagy
Article
Catalysis
Inorganic Chemistry
Protein Aggregates
03 medical and health sciences
lysosomes
Phagocytosis
Retinal Diseases
oxidized POS
medicine
Humans
Physical and Theoretical Chemistry
Molecular Biology
Retina
aging
Organic Chemistry
Retinal
Retinal Photoreceptor Cell Outer Segment
medicine.disease
eye diseases
Oxidative Stress
Autofluorescence
030104 developmental biology
lcsh:Biology (General)
lcsh:QD1-999
030221 ophthalmology & optometry
sense organs
diet
Subjects
Details
- ISSN :
- 14220067
- Volume :
- 21
- Database :
- OpenAIRE
- Journal :
- International Journal of Molecular Sciences
- Accession number :
- edsair.doi.dedup.....fd69d1b7ac6ada88f72476d3f5ae2a35