{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,25]],"date-time":"2026-02-25T10:58:39Z","timestamp":1772017119602,"version":"3.50.1"},"reference-count":85,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2020,9,11]],"date-time":"2020-09-11T00:00:00Z","timestamp":1599782400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100011718","name":"Macular Society","doi-asserted-by":"publisher","award":["PhD Studentship"],"award-info":[{"award-number":["PhD Studentship"]}],"id":[{"id":"10.13039\/100011718","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Gift of Sight Appeal","award":["N\/A"],"award-info":[{"award-number":["N\/A"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>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.<\/jats:p>","DOI":"10.3390\/ijms21186647","type":"journal-article","created":{"date-parts":[[2020,9,11]],"date-time":"2020-09-11T09:05:16Z","timestamp":1599815116000},"page":"6647","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["An In-Vitro Cell Model of Intracellular Protein Aggregation Provides Insights into RPE Stress Associated with Retinopathy"],"prefix":"10.3390","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0399-359X","authenticated-orcid":false,"given":"Eloise","family":"Keeling","sequence":"first","affiliation":[{"name":"Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, MP806, Tremona Road, Southampton SO16 6YD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annabelle J.","family":"Culling","sequence":"additional","affiliation":[{"name":"Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, MP806, Tremona Road, Southampton SO16 6YD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David A.","family":"Johnston","sequence":"additional","affiliation":[{"name":"Biomedical Imaging Unit, University of Southampton, MP12, Tremona Road, Southampton SO16 6YD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1856-3982","authenticated-orcid":false,"given":"David S.","family":"Chatelet","sequence":"additional","affiliation":[{"name":"Biomedical Imaging Unit, University of Southampton, MP12, Tremona Road, Southampton SO16 6YD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anton","family":"Page","sequence":"additional","affiliation":[{"name":"Biomedical Imaging Unit, University of Southampton, MP12, Tremona Road, Southampton SO16 6YD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5169-0561","authenticated-orcid":false,"given":"David A.","family":"Tumbarello","sequence":"additional","affiliation":[{"name":"Biological Sciences, Faculty of Environmental and Life Sciences, Life Sciences Building 85, University of Southampton, Highfield Campus, Southampton SO17 1BJ, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5541-4305","authenticated-orcid":false,"given":"Andrew J.","family":"Lotery","sequence":"additional","affiliation":[{"name":"Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, MP806, Tremona Road, Southampton SO16 6YD, UK"},{"name":"Eye Unit, University Hospital Southampton NHS Foundation Trust, Southampton SO16 6YD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1027-6938","authenticated-orcid":false,"given":"J. Arjuna","family":"Ratnayaka","sequence":"additional","affiliation":[{"name":"Clinical and Experimental Sciences, Faculty of Medicine, University of Southampton, MP806, Tremona Road, Southampton SO16 6YD, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,9,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"413","DOI":"10.2147\/OPTH.S2151","article-title":"Age and disease-related structural changes in the retinal pigment epithelium","volume":"2","author":"Bonilha","year":"2008","journal-title":"Clin. Ophthalmol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1016\/j.mam.2012.04.005","article-title":"Understanding age-related macular degeneration (AMD): Relationships between the photoreceptor\/retinal pigment epithelium\/Bruch\u2019s membrane\/choriocapillaris complex","volume":"33","author":"Bhutto","year":"2012","journal-title":"Mol. Asp. Med."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1136\/jmg.40.9.641","article-title":"The genetics of inherited macular dystrophies","volume":"40","author":"Michaelides","year":"2003","journal-title":"J. Med. Genet."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Keeling, E., Lotery, A.J., Tumbarello, D.A., and Ratnayaka, J.A. (2018). Impaired Cargo Clearance in the Retinal Pigment Epithelium (RPE) Underlies Irreversible Blinding Diseases. Cells, 7.","DOI":"10.20944\/preprints201801.0269.v1"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1083\/jcb.49.2.303","article-title":"The renewal of rod and cone outer segments in the rhesus monkey","volume":"49","author":"Young","year":"1971","journal-title":"J. Cell Biol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.preteyeres.2015.09.002","article-title":"Defects in retinal pigment epithelial cell proteolysis and the pathology associated with age-related macular degeneration","volume":"51","author":"Ferrington","year":"2016","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1374","DOI":"10.2741\/e198","article-title":"Mechanisms of protein aggregation in the retinal pigment epithelial cells","volume":"2","author":"Kaarniranta","year":"2010","journal-title":"Front. Biosci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.preteyeres.2011.12.001","article-title":"The bisretinoids of retinal pigment epithelium","volume":"31","author":"Sparrow","year":"2012","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_9","first-page":"195","article-title":"Aging human RPE: Morphometric analysis of macular, equatorial, and peripheral cells","volume":"25","author":"Hilderbrand","year":"1984","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"256","DOI":"10.1016\/S0891-5849(01)00582-2","article-title":"Photocytotoxicity of lipofuscin in human retinal pigment epithelial cells","volume":"31","author":"Davies","year":"2001","journal-title":"Free Radic. Biol. Med."},{"key":"ref_11","first-page":"3041","article-title":"Inhibition of RPE lysosomal and antioxidant activity by the age pigment lipofuscin","volume":"42","author":"Shamsi","year":"2001","journal-title":"Investig. Ophthalmol. Vis. Sci"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"5325","DOI":"10.1242\/jcs.114041","article-title":"Misregulation of autophagy and protein degradation systems in myopathies and muscular dystrophies","volume":"126","author":"Sandri","year":"2013","journal-title":"J. Cell Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1016\/S0047-6374(97)00091-2","article-title":"Preparation of artificial ceroid\/lipofuscin by UV-oxidation of subcellular organelles","volume":"99","author":"Nilsson","year":"1997","journal-title":"Mech. Ageing Dev."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"753","DOI":"10.1016\/j.exer.2004.06.025","article-title":"Transmission of light to the aging human retina: Possible implications for age related macular degeneration","volume":"79","author":"Dillon","year":"2004","journal-title":"Exp. Eye Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1167\/iovs.61.5.35","article-title":"Autofluorescent Granules of the Human Retinal Pigment Epithelium: Phenotypes, Intracellular Distribution, and Age-Related Topography","volume":"61","author":"Bermond","year":"2020","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1096\/fj.03-0289fje","article-title":"Inhibition of the ATP-driven proton pump in RPE lysosomes by the major lipofuscin fluorophore A2-E may contribute to the pathogenesis of age-related macular degeneration","volume":"18","author":"Bergmann","year":"2004","journal-title":"FASEB J."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"983","DOI":"10.1007\/s00417-002-0558-8","article-title":"Isolation of intact lysosomes from human RPE cells and effects of A2-E on the integrity of the lysosomal and other cellular membranes","volume":"240","author":"Bergmann","year":"2002","journal-title":"Graefes Arch. Clin. Exp. Ophthalmol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"3842","DOI":"10.1073\/pnas.052025899","article-title":"The lipofuscin component A2E selectively inhibits phagolysosomal degradation of photoreceptor phospholipid by the retinal pigment epithelium","volume":"99","author":"Finnemann","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"11026","DOI":"10.1073\/pnas.0702504104","article-title":"The lipofuscin fluorophore A2E perturbs cholesterol metabolism in retinal pigment epithelial cells","volume":"104","author":"Lakkaraju","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1016\/j.exer.2009.10.014","article-title":"Lipid peroxidation products reduce lysosomal protease activities in human retinal pigment epithelial cells via two different mechanisms of action","volume":"90","author":"Krohne","year":"2010","journal-title":"Exp. Eye Res."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1016\/j.exer.2009.12.011","article-title":"Effects of lipid peroxidation products on lipofuscinogenesis and autophagy in human retinal pigment epithelial cells","volume":"90","author":"Krohne","year":"2010","journal-title":"Exp. Eye Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"17929","DOI":"10.1038\/s41598-018-36322-2","article-title":"Lipofuscin-mediated photodynamic stress induces adverse changes in nanomechanical properties of retinal pigment epithelium cells","volume":"8","author":"Wiktor","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1080\/10715762.2019.1603377","article-title":"In vitro phototoxicity of rhodopsin photobleaching products in the retinal pigment epithelium (RPE)","volume":"53","author":"Olchawa","year":"2019","journal-title":"Free Radic. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"42027","DOI":"10.1074\/jbc.M305460200","article-title":"Carboxyethylpyrrole protein adducts and autoantibodies, biomarkers for age-related macular degeneration","volume":"278","author":"Gu","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1016\/j.tice.2017.06.003","article-title":"Ex-vivo models of the Retinal Pigment Epithelium (RPE) in long-term culture faithfully recapitulate key structural and physiological features of native RPE","volume":"49","author":"Lynn","year":"2017","journal-title":"Tissue Cell"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1107","DOI":"10.12688\/f1000research.15409.1","article-title":"A convenient protocol for establishing a human cell culture model of the outer retina","volume":"7","author":"Lynn","year":"2018","journal-title":"F1000Research"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Keeling, E., Chatelet, D.S., Johnston, D.A., Page, A., Tumbarello, D.A., Lotery, A.J., and Ratnayaka, J.A. (2019). Oxidative Stress and Dysfunctional Intracellular Traffic Linked to an Unhealthy Diet Results in Impaired Cargo Transport in the Retinal Pigment Epithelium (RPE). Mol. Nutr. Food Res., e1800951.","DOI":"10.1002\/mnfr.201800951"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1111\/j.1699-0463.1996.tb00717.x","article-title":"Formation of lipofuscin in cultured retinal pigment epithelial cells exposed to pre-oxidized photoreceptor outer segments","volume":"104","author":"Wihlmark","year":"1996","journal-title":"APMIS Acta Pathol. Microbiol. Immunol. Scand."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1159\/000213743","article-title":"Accumulation of lipofuscin within retinal pigment epithelial cells results in enhanced sensitivity to photo-oxidation","volume":"41","author":"Brunk","year":"1995","journal-title":"Gerontology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4351","DOI":"10.1167\/iovs.10-5852","article-title":"Interpretations of fundus autofluorescence from studies of the bisretinoids of the retina","volume":"51","author":"Sparrow","year":"2010","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Ratnayaka, J.A., Keeling, E., and Chatelet, D.S. (2019). Study of Intracellular Cargo Trafficking and Co-localization in the Phagosome and Autophagy-Lysosomal Pathways of Retinal Pigment Epithelium (RPE) Cells. Methods Mol. Biol.","DOI":"10.1007\/7651_2019_223"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1406","DOI":"10.1167\/iovs.11-8889","article-title":"Complement and UV-irradiated photoreceptor outer segments increase the cytokine secretion by retinal pigment epithelial cells","volume":"53","author":"Lueck","year":"2012","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1159\/000486404","article-title":"Activation of the ERK1\/2-MAPK Signaling Pathway by Complement Serum in UV-POS-Pretreated ARPE-19 Cells","volume":"239","author":"Busch","year":"2018","journal-title":"Ophthalmologica"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1016\/0076-6879(90)86141-H","article-title":"Determination of carbonyl content in oxidatively modified proteins","volume":"186","author":"Levine","year":"1990","journal-title":"Methods Enzymol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.exer.2014.01.010","article-title":"Understanding photoreceptor outer segment phagocytosis: Use and utility of RPE cells in culture","volume":"126","author":"Mazzoni","year":"2014","journal-title":"Exp. Eye Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3993","DOI":"10.1529\/biophysj.103.038422","article-title":"Automatic and quantitative measurement of protein-protein colocalization in live cells","volume":"86","author":"Costes","year":"2004","journal-title":"Biophys. J."},{"key":"ref_37","first-page":"459","article-title":"Pigment epithelial ensheathment and phagocytosis of extrafoveal cones in human retina","volume":"277","author":"Steinberg","year":"1977","journal-title":"Philos. Trans. R. Soc. Lond. Ser. B Biol. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Burgoyne, T., Lane, A., Laughlin, W.E., Cheetham, M.E., and Futter, C.E. (2018). Correlative light and immuno-electron microscopy of retinal tissue cryostat sections. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0191048"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1111\/acel.12072","article-title":"Balance between autophagic pathways preserves retinal homeostasis","volume":"12","author":"Koga","year":"2013","journal-title":"Aging Cell"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"973","DOI":"10.4161\/auto.24546","article-title":"Autophagy and heterophagy dysregulation leads to retinal pigment epithelium dysfunction and development of age-related macular degeneration","volume":"9","author":"Kaarniranta","year":"2013","journal-title":"Autophagy"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1016\/j.arr.2014.07.002","article-title":"Clearance of misfolded and aggregated proteins by aggrephagy and implications for aggregation diseases","volume":"18","author":"Hyttinen","year":"2014","journal-title":"Ageing Res. Rev."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"3616","DOI":"10.1111\/j.1582-4934.2008.00577.x","article-title":"Crosstalk between Hsp70 molecular chaperone, lysosomes and proteasomes in autophagy-mediated proteolysis in human retinal pigment epithelial cells","volume":"13","author":"Ryhanen","year":"2009","journal-title":"J. Cell Mol. Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"128","DOI":"10.3389\/fcell.2018.00128","article-title":"Crosstalk Between Mammalian Autophagy and the Ubiquitin-Proteasome System","volume":"6","author":"Kocaturk","year":"2018","journal-title":"Front. Cell Dev. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Blasiak, J., Pawlowska, E., Szczepanska, J., and Kaarniranta, K. (2019). Interplay between Autophagy and the Ubiquitin-Proteasome System and Its Role in the Pathogenesis of Age-Related Macular Degeneration. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20010210"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Margulis, B., Tsimokha, A., Zubova, S., and Guzhova, I. (2020). Molecular Chaperones and Proteolytic Machineries Regulate Protein Homeostasis In Aging Cells. Cells, 9.","DOI":"10.3390\/cells9051308"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Wang, A.L., Lukas, T.J., Yuan, M., Du, N., Tso, M.O., and Neufeld, A.H. (2009). Autophagy and exosomes in the aged retinal pigment epithelium: Possible relevance to drusen formation and age-related macular degeneration. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0004160"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1989","DOI":"10.4161\/auto.36184","article-title":"Dysregulated autophagy in the RPE is associated with increased susceptibility to oxidative stress and AMD","volume":"10","author":"Mitter","year":"2014","journal-title":"Autophagy"},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Viiri, J., Amadio, M., Marchesi, N., Hyttinen, J.M., Kivinen, N., Sironen, R., Rilla, K., Akhtar, S., Provenzani, A., and D\u2019Agostino, V.G. (2013). Autophagy activation clears ELAVL1\/HuR-mediated accumulation of SQSTM1\/p62 during proteasomal inhibition in human retinal pigment epithelial cells. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0069563"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1136\/bjo.78.2.125","article-title":"Regional variation and age-related changes of lysosomal enzymes in the human retinal pigment epithelium","volume":"78","author":"Boulton","year":"1994","journal-title":"Br. J. Ophthalmol."},{"key":"ref_50","first-page":"1138","article-title":"Lysosomal enzyme cytochemistry of human RPE, Bruch\u2019s membrane and drusen","volume":"28","author":"Gao","year":"1987","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"e2537","DOI":"10.1038\/cddis.2016.453","article-title":"Dysfunctional autophagy in RPE, a contributing factor in age-related macular degeneration","volume":"8","author":"Golestaneh","year":"2017","journal-title":"Cell Death Dis."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Kozhevnikova, O.S., Telegina, D.V., Tyumentsev, M.A., and Kolosova, N.G. (2019). Disruptions of Autophagy in the Rat Retina with Age During the Development of Age-Related-Macular-Degeneration-like Retinopathy. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20194804"},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Qi, X., Mitter, S.K., Yan, Y., Busik, J.V., Grant, M.B., and Boulton, M.E. (2020). Diurnal Rhythmicity of Autophagy Is Impaired in the Diabetic Retina. Cells, 9.","DOI":"10.3390\/cells9040905"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"20639","DOI":"10.1038\/srep20639","article-title":"Impairing autophagy in retinal pigment epithelium leads to inflammasome activation and enhanced macrophage-mediated angiogenesis","volume":"6","author":"Liu","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"480","DOI":"10.4161\/auto.27292","article-title":"Lysosomal-mediated waste clearance in retinal pigment epithelial cells is regulated by CRYBA1\/betaA3\/A1-crystallin via V-ATPase-MTORC1 signaling","volume":"10","author":"Valapala","year":"2014","journal-title":"Autophagy"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"13094","DOI":"10.1073\/pnas.2000339117","article-title":"Altered photoreceptor metabolism in mouse causes late stage age-related macular degeneration-like pathologies","volume":"117","author":"Cheng","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_57","first-page":"1029","article-title":"PO2 profiles and oxygen consumption in cat retina with an occluded retinal circulation","volume":"31","author":"Alder","year":"1990","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Roehlecke, C., Schumann, U., Ader, M., Brunssen, C., Bramke, S., Morawietz, H., and Funk, R.H. (2013). Stress reaction in outer segments of photoreceptors after blue light irradiation. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0071570"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/j.biochi.2016.03.016","article-title":"Impairment of extramitochondrial oxidative phosphorylation in mouse rod outer segments by blue light irradiation","volume":"125","author":"Calzia","year":"2016","journal-title":"Biochimie"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1021\/acs.jproteome.7b00741","article-title":"Proteome of Bovine Mitochondria and Rod Outer Segment Disks: Commonalities and Differences","volume":"17","author":"Bruschi","year":"2018","journal-title":"J. Proteome Res."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.freeradbiomed.2019.01.040","article-title":"Corrigendum to \u201cModulation of the rod outer segment aerobic metabolism diminishes the production of radicals due to light absorption\u201d","volume":"136","author":"Calzia","year":"2019","journal-title":"Free Radic. Biol. Med."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"1775","DOI":"10.1167\/iovs.02-0742","article-title":"Induction of angiogenic cytokine expression in cultured RPE by ingestion of oxidized photoreceptor outer segments","volume":"44","author":"Higgins","year":"2003","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"635","DOI":"10.1016\/j.exer.2006.11.015","article-title":"Synthesis of complement factor H by retinal pigment epithelial cells is down-regulated by oxidized photoreceptor outer segments","volume":"84","author":"Chen","year":"2007","journal-title":"Exp. Eye Res."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"2052","DOI":"10.18632\/aging.101299","article-title":"PKC\u03b6-dependent upregulation of p27kip1 contributes to oxidative stress induced retinal pigment epithelial cell multinucleation","volume":"9","author":"Rajapakse","year":"2017","journal-title":"Aging"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"1380","DOI":"10.18632\/aging.101474","article-title":"The expression of C1 inhibitor (C1INH) in macrophages is upregulated by retinal pigment epithelial cells\u2014implication in subretinal immune privilege in the aging eye","volume":"10","author":"Luo","year":"2018","journal-title":"Aging"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"851","DOI":"10.1080\/02713689808951268","article-title":"Lipofuscin accumulation in cultured retinal pigment epithelial cells reduces their phagocytic capacity","volume":"17","author":"Sundelin","year":"1998","journal-title":"Curr. Eye Res."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"6926485","DOI":"10.1155\/2017\/6926485","article-title":"Zinc Protects Oxidative Stress-Induced RPE Death by Reducing Mitochondrial Damage and Preventing Lysosome Rupture","volume":"2017","author":"Rajapakse","year":"2017","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"1397","DOI":"10.1074\/mcp.M700525-MCP200","article-title":"Retinal pigment epithelium lipofuscin proteomics","volume":"7","author":"Ng","year":"2008","journal-title":"Mol. Cell. Proteom. MCP"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"9014","DOI":"10.1073\/pnas.1805039115","article-title":"Aberrant early endosome biogenesis mediates complement activation in the retinal pigment epithelium in models of macular degeneration","volume":"115","author":"Kaur","year":"2018","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1016\/S0002-9440(10)64538-5","article-title":"Endocytic pathway abnormalities precede amyloid beta deposition in sporadic Alzheimer\u2019s disease and Down syndrome: Differential effects of APOE genotype and presenilin mutations","volume":"157","author":"Cataldo","year":"2000","journal-title":"Am. J. Pathol."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"1263","DOI":"10.1016\/j.neurobiolaging.2004.02.027","article-title":"Abeta localization in abnormal endosomes: Association with earliest Abeta elevations in AD and Down syndrome","volume":"25","author":"Cataldo","year":"2004","journal-title":"Neurobiol. Aging"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/15548627.2015.1100356","article-title":"Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)","volume":"12","author":"Klionsky","year":"2016","journal-title":"Autophagy"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1016\/j.ophtha.2016.12.002","article-title":"Imaging Protocols in Clinical Studies in Advanced Age-Related Macular Degeneration: Recommendations from Classification of Atrophy Consensus Meetings","volume":"124","author":"Holz","year":"2017","journal-title":"Ophthalmology"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1007\/s004170050209","article-title":"Patterns of increased in vivo fundus autofluorescence in the junctional zone of geographic atrophy of the retinal pigment epithelium associated with age-related macular degeneration","volume":"237","author":"Holz","year":"1999","journal-title":"Graefes Arch. Clin. Exp. Ophthalmol."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1016\/j.ajo.2006.11.041","article-title":"Progression of geographic atrophy and impact of fundus autofluorescence patterns in age-related macular degeneration","volume":"143","author":"Holz","year":"2007","journal-title":"Am. J. Ophthalmol."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"2481","DOI":"10.1167\/iovs.19-26949","article-title":"Quantifying Retinal Pigment Epithelium Dysmorphia and Loss of Histologic Autofluorescence in Age-Related Macular Degeneration","volume":"60","author":"Gambril","year":"2019","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1001\/jamaophthalmol.2016.1475","article-title":"Quantitative Fundus Autofluorescence in Early and Intermediate Age-Related Macular Degeneration","volume":"134","author":"Gliem","year":"2016","journal-title":"JAMA Ophthalmol."},{"key":"ref_78","first-page":"S34","article-title":"Quantitative Fundus Autofluorescence in Non-Neovascular Age-Related Macular Degeneration","volume":"49","author":"Yokoyama","year":"2018","journal-title":"Ophthalmic Surg. Lasers Imaging Retin."},{"key":"ref_79","first-page":"718","article-title":"In vivo fluorescence of the ocular fundus exhibits retinal pigment epithelium lipofuscin characteristics","volume":"36","author":"Delori","year":"1995","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1016\/j.euroneuro.2013.03.007","article-title":"Proteinopathies, a core concept for understanding and ultimately treating degenerative disorders?","volume":"25","author":"Bayer","year":"2015","journal-title":"Eur. Neuropsychopharmacol. J. Eur. Coll. Neuropsychopharmacol."},{"key":"ref_81","doi-asserted-by":"crossref","unstructured":"Datta, S., Cano, M., Ebrahimi, K., Wang, L., and Handa, J.T. (2017). The impact of oxidative stress and inflammation on RPE degeneration in non-neovascular AMD. Prog. Retin. Eye Res.","DOI":"10.1016\/j.preteyeres.2017.03.002"},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"863","DOI":"10.4103\/1673-5374.268972","article-title":"Challenges in studying geographic atrophy (GA) age-related macular degeneration: The potential of a new mouse model with GA-like features","volume":"15","author":"Ratnayaka","year":"2020","journal-title":"Neural Regen. Res."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1006\/exer.1996.0020","article-title":"ARPE-19, a human retinal pigment epithelial cell line with differentiated properties","volume":"62","author":"Dunn","year":"1996","journal-title":"Exp. Eye Res."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"907","DOI":"10.1016\/S0014-4835(87)80105-7","article-title":"Kinetic studies of rod outer segment binding and ingestion by cultured rat RPE cells","volume":"45","author":"Hall","year":"1987","journal-title":"Exp. Eye Res."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/18\/6647\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:08:58Z","timestamp":1760177338000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/21\/18\/6647"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,11]]},"references-count":85,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["ijms21186647"],"URL":"https:\/\/doi.org\/10.3390\/ijms21186647","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,9,11]]}}}