Single Cell Analysis of Reversibility of the Cell Death Program in Ethanol-Treated Neuronal PC12 Cells
Neurodegenerative diseases are generally characterized clinically by the selective loss of a distinct subset of neurons and a slow progressive course. Mounting evidence in vivo indicates that large numbers of neurons pass through a long period of injury and dysfunction before the actual death of the...
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MDPI AG
2022-02-01
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| Series: | International Journal of Molecular Sciences |
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| author | Wenting You Tos T. J. M. Berendschot Kèvin Knoops Marc A. M. J. van Zandvoort Carroll A. B. Webers Chris P. M. Reutelingsperger Theo G. M. F. Gorgels |
| author_facet | Wenting You Tos T. J. M. Berendschot Kèvin Knoops Marc A. M. J. van Zandvoort Carroll A. B. Webers Chris P. M. Reutelingsperger Theo G. M. F. Gorgels |
| author_sort | Wenting You |
| collection | DOAJ |
| description | Neurodegenerative diseases are generally characterized clinically by the selective loss of a distinct subset of neurons and a slow progressive course. Mounting evidence in vivo indicates that large numbers of neurons pass through a long period of injury and dysfunction before the actual death of the cells. Whether these dying neurons can be rescued and return to a normal, functional state is uncertain. In the present study, we explored the reversibility of the neuronal cell death pathway at various stages by monitoring the dynamics of single cells with high-resolution live-cell spinning disk confocal microscopy in an in vitro neuronal cell death model. We exposed differentiated neuronal PC12 cells to ethanol as our cell death model. Results showed that exposure to 5% ethanol for 24 h induced cell death in >70% of the cells. Ethanol treatment for 3 h already induced cellular changes and damage such as reactive oxygen species generation, elevation of intracellular Ca<sup>2+</sup> level, phosphatidylserine exposure, nuclear shrinkage, DNA damage, mitochondrial fragmentation and membrane potential loss, and retraction of neurites. These phenomena are often associated with programmed cell death. Importantly, after removing ethanol and further culturing these damaged cells in fresh culture medium, cells recovered from all these cell injuries and generated new neurites. Moreover, results indicated that this recovery was not dependent on exogenous NGF and other growth factors in the cell culture medium. Overall, our results suggest that targeting dying neurons can be an effective therapeutic strategy in neurodegenerative diseases. |
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| language | English |
| last_indexed | 2024-03-09T20:36:33Z |
| publishDate | 2022-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-0ce9342e62af4b17bd53e441487b99fb2023-11-23T23:07:29ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-02-01235265010.3390/ijms23052650Single Cell Analysis of Reversibility of the Cell Death Program in Ethanol-Treated Neuronal PC12 CellsWenting You0Tos T. J. M. Berendschot1Kèvin Knoops2Marc A. M. J. van Zandvoort3Carroll A. B. Webers4Chris P. M. Reutelingsperger5Theo G. M. F. Gorgels6University Eye Clinic Maastricht UMC+, Maastricht University Medical Center+, 6229 HX Maastricht, The NetherlandsUniversity Eye Clinic Maastricht UMC+, Maastricht University Medical Center+, 6229 HX Maastricht, The NetherlandsThe Maastricht Multimodal Molecular Imaging Institute, Maastricht University, 6229 ER Maastricht, The NetherlandsDepartment of Molecular Cell Biology, CARIM School for Cardiovascular Disease, Maastricht University, 6229 ER Maastricht, The NetherlandsUniversity Eye Clinic Maastricht UMC+, Maastricht University Medical Center+, 6229 HX Maastricht, The NetherlandsDepartment of Biochemistry, CARIM School for Cardiovascular Disease, Maastricht University, 6229 ER Maastricht, The NetherlandsUniversity Eye Clinic Maastricht UMC+, Maastricht University Medical Center+, 6229 HX Maastricht, The NetherlandsNeurodegenerative diseases are generally characterized clinically by the selective loss of a distinct subset of neurons and a slow progressive course. Mounting evidence in vivo indicates that large numbers of neurons pass through a long period of injury and dysfunction before the actual death of the cells. Whether these dying neurons can be rescued and return to a normal, functional state is uncertain. In the present study, we explored the reversibility of the neuronal cell death pathway at various stages by monitoring the dynamics of single cells with high-resolution live-cell spinning disk confocal microscopy in an in vitro neuronal cell death model. We exposed differentiated neuronal PC12 cells to ethanol as our cell death model. Results showed that exposure to 5% ethanol for 24 h induced cell death in >70% of the cells. Ethanol treatment for 3 h already induced cellular changes and damage such as reactive oxygen species generation, elevation of intracellular Ca<sup>2+</sup> level, phosphatidylserine exposure, nuclear shrinkage, DNA damage, mitochondrial fragmentation and membrane potential loss, and retraction of neurites. These phenomena are often associated with programmed cell death. Importantly, after removing ethanol and further culturing these damaged cells in fresh culture medium, cells recovered from all these cell injuries and generated new neurites. Moreover, results indicated that this recovery was not dependent on exogenous NGF and other growth factors in the cell culture medium. Overall, our results suggest that targeting dying neurons can be an effective therapeutic strategy in neurodegenerative diseases.https://www.mdpi.com/1422-0067/23/5/2650cell death programneurite retractionneurite regenerationnuclear shrinkagemitochondrial fragmentationPS exposure |
| spellingShingle | Wenting You Tos T. J. M. Berendschot Kèvin Knoops Marc A. M. J. van Zandvoort Carroll A. B. Webers Chris P. M. Reutelingsperger Theo G. M. F. Gorgels Single Cell Analysis of Reversibility of the Cell Death Program in Ethanol-Treated Neuronal PC12 Cells International Journal of Molecular Sciences cell death program neurite retraction neurite regeneration nuclear shrinkage mitochondrial fragmentation PS exposure |
| title | Single Cell Analysis of Reversibility of the Cell Death Program in Ethanol-Treated Neuronal PC12 Cells |
| title_full | Single Cell Analysis of Reversibility of the Cell Death Program in Ethanol-Treated Neuronal PC12 Cells |
| title_fullStr | Single Cell Analysis of Reversibility of the Cell Death Program in Ethanol-Treated Neuronal PC12 Cells |
| title_full_unstemmed | Single Cell Analysis of Reversibility of the Cell Death Program in Ethanol-Treated Neuronal PC12 Cells |
| title_short | Single Cell Analysis of Reversibility of the Cell Death Program in Ethanol-Treated Neuronal PC12 Cells |
| title_sort | single cell analysis of reversibility of the cell death program in ethanol treated neuronal pc12 cells |
| topic | cell death program neurite retraction neurite regeneration nuclear shrinkage mitochondrial fragmentation PS exposure |
| url | https://www.mdpi.com/1422-0067/23/5/2650 |
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