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An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations

PURPOSE: was to create an in vitro model of human retinal detachment (RD) to study the mechanisms of photoreceptor death. METHODS: Human retinas were obtained through eye globe donations for research purposes and cultivated as explants. Cell death was investigated in retinas with (control) and witho...

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Autores principales: Potic, Jelena, Mbefo, Martial, Berger, Adeline, Nicolas, Michael, Wanner, Dana, Kostic, Corinne, Matet, Alexandre, Behar-Cohen, Francine, Moulin, Alexandre, Arsenijevic, Yvan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7726250/
https://www.ncbi.nlm.nih.gov/pubmed/33324144
http://dx.doi.org/10.3389/fnins.2020.571293
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author Potic, Jelena
Mbefo, Martial
Berger, Adeline
Nicolas, Michael
Wanner, Dana
Kostic, Corinne
Matet, Alexandre
Behar-Cohen, Francine
Moulin, Alexandre
Arsenijevic, Yvan
author_facet Potic, Jelena
Mbefo, Martial
Berger, Adeline
Nicolas, Michael
Wanner, Dana
Kostic, Corinne
Matet, Alexandre
Behar-Cohen, Francine
Moulin, Alexandre
Arsenijevic, Yvan
author_sort Potic, Jelena
collection PubMed
description PURPOSE: was to create an in vitro model of human retinal detachment (RD) to study the mechanisms of photoreceptor death. METHODS: Human 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. RESULTS: The 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. CONCLUSION: AIF 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.
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spelling pubmed-77262502020-12-14 An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations Potic, Jelena Mbefo, Martial Berger, Adeline Nicolas, Michael Wanner, Dana Kostic, Corinne Matet, Alexandre Behar-Cohen, Francine Moulin, Alexandre Arsenijevic, Yvan Front Neurosci Neuroscience PURPOSE: was to create an in vitro model of human retinal detachment (RD) to study the mechanisms of photoreceptor death. METHODS: Human 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. RESULTS: The 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. CONCLUSION: AIF 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. Frontiers Media S.A. 2020-11-26 /pmc/articles/PMC7726250/ /pubmed/33324144 http://dx.doi.org/10.3389/fnins.2020.571293 Text en Copyright © 2020 Potic, Mbefo, Berger, Nicolas, Wanner, Kostic, Matet, Behar-Cohen, Moulin and Arsenijevic. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Neuroscience
Potic, Jelena
Mbefo, Martial
Berger, Adeline
Nicolas, Michael
Wanner, Dana
Kostic, Corinne
Matet, Alexandre
Behar-Cohen, Francine
Moulin, Alexandre
Arsenijevic, Yvan
An in vitro Model of Human Retinal Detachment Reveals Successive Death Pathway Activations
title 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_short 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
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7726250/
https://www.ncbi.nlm.nih.gov/pubmed/33324144
http://dx.doi.org/10.3389/fnins.2020.571293
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