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An automated fluorescence videomicroscopy assay for the detection of mitotic catastrophe

Mitotic catastrophe can be defined as a cell death mode that occurs during or shortly after a prolonged/aberrant mitosis, and can show apoptotic or necrotic features. However, conventional procedures for the detection of apoptosis or necrosis, including biochemical bulk assays and cytofluorometric t...

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Autores principales: Rello-Varona, S, Kepp, O, Vitale, I, Michaud, M, Senovilla, L, Jemaà, M, Joza, N, Galluzzi, L, Castedo, M, Kroemer, G
Formato: Texto
Lenguaje:English
Publicado: Nature Publishing Group 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3032329/
https://www.ncbi.nlm.nih.gov/pubmed/21364633
http://dx.doi.org/10.1038/cddis.2010.6
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author Rello-Varona, S
Kepp, O
Vitale, I
Michaud, M
Senovilla, L
Jemaà, M
Joza, N
Galluzzi, L
Castedo, M
Kroemer, G
author_facet Rello-Varona, S
Kepp, O
Vitale, I
Michaud, M
Senovilla, L
Jemaà, M
Joza, N
Galluzzi, L
Castedo, M
Kroemer, G
author_sort Rello-Varona, S
collection PubMed
description Mitotic catastrophe can be defined as a cell death mode that occurs during or shortly after a prolonged/aberrant mitosis, and can show apoptotic or necrotic features. However, conventional procedures for the detection of apoptosis or necrosis, including biochemical bulk assays and cytofluorometric techniques, cannot discriminate among pre-mitotic, mitotic and post-mitotic death, and hence are inappropriate to monitor mitotic catastrophe. To address this issue, we generated isogenic human colon carcinoma cell lines that differ in ploidy and p53 status, yet express similar amounts of fluorescent biosensors that allow for the visualization of chromatin (histone H2B coupled to green fluorescent protein (GFP)) and centrosomes (centrin coupled to the Discosoma striata red fluorescent protein (DsRed)). By combining high-resolution fluorescence videomicroscopy and automated image analysis, we established protocols and settings for the simultaneous assessment of ploidy, mitosis, centrosome number and cell death (which in our model system occurs mainly by apoptosis). Time-lapse videomicroscopy showed that this approach can be used for the high-throughput detection of mitotic catastrophe induced by three mechanistically distinct anti-mitotic agents (dimethylenastron (DIMEN), nocodazole (NDZ) and paclitaxel (PTX)), and – in this context – revealed an important role of p53 in the control of centrosome number.
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spelling pubmed-30323292011-02-24 An automated fluorescence videomicroscopy assay for the detection of mitotic catastrophe Rello-Varona, S Kepp, O Vitale, I Michaud, M Senovilla, L Jemaà, M Joza, N Galluzzi, L Castedo, M Kroemer, G Cell Death Dis Original Article Mitotic catastrophe can be defined as a cell death mode that occurs during or shortly after a prolonged/aberrant mitosis, and can show apoptotic or necrotic features. However, conventional procedures for the detection of apoptosis or necrosis, including biochemical bulk assays and cytofluorometric techniques, cannot discriminate among pre-mitotic, mitotic and post-mitotic death, and hence are inappropriate to monitor mitotic catastrophe. To address this issue, we generated isogenic human colon carcinoma cell lines that differ in ploidy and p53 status, yet express similar amounts of fluorescent biosensors that allow for the visualization of chromatin (histone H2B coupled to green fluorescent protein (GFP)) and centrosomes (centrin coupled to the Discosoma striata red fluorescent protein (DsRed)). By combining high-resolution fluorescence videomicroscopy and automated image analysis, we established protocols and settings for the simultaneous assessment of ploidy, mitosis, centrosome number and cell death (which in our model system occurs mainly by apoptosis). Time-lapse videomicroscopy showed that this approach can be used for the high-throughput detection of mitotic catastrophe induced by three mechanistically distinct anti-mitotic agents (dimethylenastron (DIMEN), nocodazole (NDZ) and paclitaxel (PTX)), and – in this context – revealed an important role of p53 in the control of centrosome number. Nature Publishing Group 2010-02 2010-02-18 /pmc/articles/PMC3032329/ /pubmed/21364633 http://dx.doi.org/10.1038/cddis.2010.6 Text en Copyright © 2010 Macmillan Publishers Limited http://creativecommons.org/licenses/by-nc-nd/3.0/ This article is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Original Article
Rello-Varona, S
Kepp, O
Vitale, I
Michaud, M
Senovilla, L
Jemaà, M
Joza, N
Galluzzi, L
Castedo, M
Kroemer, G
An automated fluorescence videomicroscopy assay for the detection of mitotic catastrophe
title An automated fluorescence videomicroscopy assay for the detection of mitotic catastrophe
title_full An automated fluorescence videomicroscopy assay for the detection of mitotic catastrophe
title_fullStr An automated fluorescence videomicroscopy assay for the detection of mitotic catastrophe
title_full_unstemmed An automated fluorescence videomicroscopy assay for the detection of mitotic catastrophe
title_short An automated fluorescence videomicroscopy assay for the detection of mitotic catastrophe
title_sort automated fluorescence videomicroscopy assay for the detection of mitotic catastrophe
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3032329/
https://www.ncbi.nlm.nih.gov/pubmed/21364633
http://dx.doi.org/10.1038/cddis.2010.6
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