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Relationships of diverse apoptotic death process patterns to mitochondrial membrane potential (Δψ(m)) evaluated by three-parameter flow cytometric analysis
Recently, it has been proposed that novel methodologies are needed to re-evaluate apoptotic cell death, as studies of apoptosis have shown it to be a complex process. Since mitochondria are key regulators in cell death pathways, we developed a simultaneous 3-parameter flow cytometric analysis that i...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Netherlands
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3536873/ https://www.ncbi.nlm.nih.gov/pubmed/22669602 http://dx.doi.org/10.1007/s10616-012-9455-0 |
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author | Suzuki, Yuhgi Hasegawa, Hiroo Tsuji, Tomohiro Tsuruda, Kazuto Sasaki, Daisuke Ishihara, Kaori Nagai, Kazuhiro Yanagihara, Katsunori Yamada, Yasuaki Kamihira, Shimeru |
author_facet | Suzuki, Yuhgi Hasegawa, Hiroo Tsuji, Tomohiro Tsuruda, Kazuto Sasaki, Daisuke Ishihara, Kaori Nagai, Kazuhiro Yanagihara, Katsunori Yamada, Yasuaki Kamihira, Shimeru |
author_sort | Suzuki, Yuhgi |
collection | PubMed |
description | Recently, it has been proposed that novel methodologies are needed to re-evaluate apoptotic cell death, as studies of apoptosis have shown it to be a complex process. Since mitochondria are key regulators in cell death pathways, we developed a simultaneous 3-parameter flow cytometric analysis that incorporates the change in mitochondrial membrane potential (Δψ(m)) in an Annexin-V [for phosphatidyl-serine (PS)] and propidium iodide (PI) assay system (3 parameters with 4 colours), and evaluated the apoptotic process using various haematological malignant cell lines and death triggers. The present method enabled visualization of cell composition during apoptosis and captured complicated molecular events. For example, apoptotic cells that lost Δψ(m) did not always externalize PS, while some late apoptotic cells had polarized Δψ(m). The findings of unchanged PS-externalization and aberrant cell death suggest that there is no relationship of PS externalization and apoptosis with an unknown apoptotic mechanism. Based on PS-externalization, sensitivity to staurosporine, and the combination of cell lines and triggers, the apoptotic process was classified into 2 types. Importantly, most of our findings could not be observed by PS–PI and Δψ(m) assays when independently performed. Our method may be useful for examining mitochondrial-related apoptosis and death signalling pathways, as well as screening novel apoptosis-inducing cancer drugs. |
format | Online Article Text |
id | pubmed-3536873 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Springer Netherlands |
record_format | MEDLINE/PubMed |
spelling | pubmed-35368732013-01-04 Relationships of diverse apoptotic death process patterns to mitochondrial membrane potential (Δψ(m)) evaluated by three-parameter flow cytometric analysis Suzuki, Yuhgi Hasegawa, Hiroo Tsuji, Tomohiro Tsuruda, Kazuto Sasaki, Daisuke Ishihara, Kaori Nagai, Kazuhiro Yanagihara, Katsunori Yamada, Yasuaki Kamihira, Shimeru Cytotechnology Original Research Recently, it has been proposed that novel methodologies are needed to re-evaluate apoptotic cell death, as studies of apoptosis have shown it to be a complex process. Since mitochondria are key regulators in cell death pathways, we developed a simultaneous 3-parameter flow cytometric analysis that incorporates the change in mitochondrial membrane potential (Δψ(m)) in an Annexin-V [for phosphatidyl-serine (PS)] and propidium iodide (PI) assay system (3 parameters with 4 colours), and evaluated the apoptotic process using various haematological malignant cell lines and death triggers. The present method enabled visualization of cell composition during apoptosis and captured complicated molecular events. For example, apoptotic cells that lost Δψ(m) did not always externalize PS, while some late apoptotic cells had polarized Δψ(m). The findings of unchanged PS-externalization and aberrant cell death suggest that there is no relationship of PS externalization and apoptosis with an unknown apoptotic mechanism. Based on PS-externalization, sensitivity to staurosporine, and the combination of cell lines and triggers, the apoptotic process was classified into 2 types. Importantly, most of our findings could not be observed by PS–PI and Δψ(m) assays when independently performed. Our method may be useful for examining mitochondrial-related apoptosis and death signalling pathways, as well as screening novel apoptosis-inducing cancer drugs. Springer Netherlands 2012-06-06 2013-01 /pmc/articles/PMC3536873/ /pubmed/22669602 http://dx.doi.org/10.1007/s10616-012-9455-0 Text en © The Author(s) 2012 https://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. |
spellingShingle | Original Research Suzuki, Yuhgi Hasegawa, Hiroo Tsuji, Tomohiro Tsuruda, Kazuto Sasaki, Daisuke Ishihara, Kaori Nagai, Kazuhiro Yanagihara, Katsunori Yamada, Yasuaki Kamihira, Shimeru Relationships of diverse apoptotic death process patterns to mitochondrial membrane potential (Δψ(m)) evaluated by three-parameter flow cytometric analysis |
title | Relationships of diverse apoptotic death process patterns to mitochondrial membrane potential (Δψ(m)) evaluated by three-parameter flow cytometric analysis |
title_full | Relationships of diverse apoptotic death process patterns to mitochondrial membrane potential (Δψ(m)) evaluated by three-parameter flow cytometric analysis |
title_fullStr | Relationships of diverse apoptotic death process patterns to mitochondrial membrane potential (Δψ(m)) evaluated by three-parameter flow cytometric analysis |
title_full_unstemmed | Relationships of diverse apoptotic death process patterns to mitochondrial membrane potential (Δψ(m)) evaluated by three-parameter flow cytometric analysis |
title_short | Relationships of diverse apoptotic death process patterns to mitochondrial membrane potential (Δψ(m)) evaluated by three-parameter flow cytometric analysis |
title_sort | relationships of diverse apoptotic death process patterns to mitochondrial membrane potential (δψ(m)) evaluated by three-parameter flow cytometric analysis |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3536873/ https://www.ncbi.nlm.nih.gov/pubmed/22669602 http://dx.doi.org/10.1007/s10616-012-9455-0 |
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