An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations
Purposewas to create an in vitro model of human retinal detachment (RD) to study the mechanisms of photoreceptor death.MethodsHuman retinas were obtained through eye globe donations for research purposes and cultivated as explants. Cell death was investigated in retinas with (control) and without re...
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doaj-ec20cdd3938f4fa49562af439ca068eb2020-12-08T08:42:14ZengFrontiers Media S.A.Frontiers in Neuroscience1662-453X2020-11-011410.3389/fnins.2020.571293571293An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway ActivationsJelena Potic0Jelena Potic1Jelena Potic2Martial Mbefo3Adeline Berger4Michael Nicolas5Dana Wanner6Corinne Kostic7Alexandre Matet8Alexandre Matet9Francine Behar-Cohen10Francine Behar-Cohen11Alexandre Moulin12Yvan Arsenijevic13Department of Ophthalmology, University of Lausanne, Jules-Gonin Eye Hospital, Fondation Asile des Aveugles, Lausanne, SwitzerlandClinic for Eye Diseases, Clinical Center of Serbia, Belgrade, SerbiaDepartment of Ophthalmology, Faculty of Medicine, University of Belgrade, Belgrade, SerbiaDepartment of Ophthalmology, University of Lausanne, Jules-Gonin Eye Hospital, Fondation Asile des Aveugles, Lausanne, SwitzerlandDepartment of Ophthalmology, University of Lausanne, Jules-Gonin Eye Hospital, Fondation Asile des Aveugles, Lausanne, SwitzerlandDepartment of Ophthalmology, University of Lausanne, Jules-Gonin Eye Hospital, Fondation Asile des Aveugles, Lausanne, SwitzerlandDepartment of Ophthalmology, University of Lausanne, Jules-Gonin Eye Hospital, Fondation Asile des Aveugles, Lausanne, SwitzerlandDepartment of Ophthalmology, University of Lausanne, Jules-Gonin Eye Hospital, Fondation Asile des Aveugles, Lausanne, SwitzerlandDepartment of Ophthalmology, University of Lausanne, Jules-Gonin Eye Hospital, Fondation Asile des Aveugles, Lausanne, SwitzerlandDepartment of Ophthalmology, Institut Curie, Université de Paris, Paris, FranceDepartment of Ophthalmology, University of Lausanne, Jules-Gonin Eye Hospital, Fondation Asile des Aveugles, Lausanne, SwitzerlandINSERM U 1138, Centre de Recherches des Cordeliers, Université Paris Descartes, Université Pierre et Marie Curie, Paris, FranceDepartment of Ophthalmology, University of Lausanne, Jules-Gonin Eye Hospital, Fondation Asile des Aveugles, Lausanne, SwitzerlandDepartment of Ophthalmology, University of Lausanne, Jules-Gonin Eye Hospital, Fondation Asile des Aveugles, Lausanne, SwitzerlandPurposewas to create an in vitro model of human retinal detachment (RD) to study the mechanisms of photoreceptor death.MethodsHuman retinas were obtained through eye globe donations for research purposes and cultivated as explants. Cell death was investigated in retinas with (control) and without retinal pigment epithelium (RPE) cells to mimic RD. Tissues were studied at different time points and immunohistological analyses for TUNEL, Cleaved caspase3, AIF, CDK4 and the epigenetic mark H3K27me3 were performed. Human and monkey eye globes with retinal detachment served as controls.ResultsThe number of TUNEL-positive cells, compared between 1 and 7 days, increased with time in both retinas with RPE (from 1.2 ± 0.46 to 8 ± 0.89, n = 4) and without RPE (from 2.6 ± 0.73 to 16.3 ± 1.27, p < 0.014). In the group without RPE, cell death peaked at day 3 (p = 0.014) and was high until day 7. Almost no Cleaved-Caspase3 signal was observed, whereas a transient augmentation at day 3 of AIF-positive cells was observed to be about 10-fold in comparison to the control group (n = 2). Few CDK4-positive cells were found in both groups, but significantly more in the RD group at day 7 (1.8 ± 0.24 vs. 4.7 ± 0.58, p = 0.014). The H3K27me3 mark increased by 7-fold after 5 days in the RD group (p = 0.014) and slightly decreased at day 7 and was also observed to be markedly increased in human and monkey detached retina samples.ConclusionAIF expression coincides with the first peak of cell death, whereas the H3K27me3 mark increases during the cell death plateau, suggesting that photoreceptor death is induced by different successive pathways after RD. This in vitro model should permit the identification of neuroprotective drugs with clinical relevance.https://www.frontiersin.org/articles/10.3389/fnins.2020.571293/fullhuman retinain vitro modelretinal detachmentphotoreceptor deathcell death pathways |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jelena Potic Jelena Potic Jelena Potic Martial Mbefo Adeline Berger Michael Nicolas Dana Wanner Corinne Kostic Alexandre Matet Alexandre Matet Francine Behar-Cohen Francine Behar-Cohen Alexandre Moulin Yvan Arsenijevic |
spellingShingle |
Jelena Potic Jelena Potic Jelena Potic Martial Mbefo Adeline Berger Michael Nicolas Dana Wanner Corinne Kostic Alexandre Matet Alexandre Matet Francine Behar-Cohen Francine Behar-Cohen Alexandre Moulin Yvan Arsenijevic An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations Frontiers in Neuroscience human retina in vitro model retinal detachment photoreceptor death cell death pathways |
author_facet |
Jelena Potic Jelena Potic Jelena Potic Martial Mbefo Adeline Berger Michael Nicolas Dana Wanner Corinne Kostic Alexandre Matet Alexandre Matet Francine Behar-Cohen Francine Behar-Cohen Alexandre Moulin Yvan Arsenijevic |
author_sort |
Jelena Potic |
title |
An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations |
title_short |
An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations |
title_full |
An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations |
title_fullStr |
An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations |
title_full_unstemmed |
An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations |
title_sort |
in vitro model of human retinal detachment reveals successive death pathway activations |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Neuroscience |
issn |
1662-453X |
publishDate |
2020-11-01 |
description |
Purposewas to create an in vitro model of human retinal detachment (RD) to study the mechanisms of photoreceptor death.MethodsHuman retinas were obtained through eye globe donations for research purposes and cultivated as explants. Cell death was investigated in retinas with (control) and without retinal pigment epithelium (RPE) cells to mimic RD. Tissues were studied at different time points and immunohistological analyses for TUNEL, Cleaved caspase3, AIF, CDK4 and the epigenetic mark H3K27me3 were performed. Human and monkey eye globes with retinal detachment served as controls.ResultsThe number of TUNEL-positive cells, compared between 1 and 7 days, increased with time in both retinas with RPE (from 1.2 ± 0.46 to 8 ± 0.89, n = 4) and without RPE (from 2.6 ± 0.73 to 16.3 ± 1.27, p < 0.014). In the group without RPE, cell death peaked at day 3 (p = 0.014) and was high until day 7. Almost no Cleaved-Caspase3 signal was observed, whereas a transient augmentation at day 3 of AIF-positive cells was observed to be about 10-fold in comparison to the control group (n = 2). Few CDK4-positive cells were found in both groups, but significantly more in the RD group at day 7 (1.8 ± 0.24 vs. 4.7 ± 0.58, p = 0.014). The H3K27me3 mark increased by 7-fold after 5 days in the RD group (p = 0.014) and slightly decreased at day 7 and was also observed to be markedly increased in human and monkey detached retina samples.ConclusionAIF expression coincides with the first peak of cell death, whereas the H3K27me3 mark increases during the cell death plateau, suggesting that photoreceptor death is induced by different successive pathways after RD. This in vitro model should permit the identification of neuroprotective drugs with clinical relevance. |
topic |
human retina in vitro model retinal detachment photoreceptor death cell death pathways |
url |
https://www.frontiersin.org/articles/10.3389/fnins.2020.571293/full |
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