Inactivation of MARCH5 prevents mitochondrial fragmentation and interferes with cell death in a neuronal cell model.
To study the impact of the mitochondrial ubiquitin ligase MARCH5 on mitochondrial morphology and induction of apoptosis using an in vitro model of neuronal precursor cells exposed to glaucoma-relevant stress conditions.RGC5 cells transfected with expression constructs for MARCH5, MARCH5(H43W), Dpr1(...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3526576?pdf=render |
| _version_ | 1819213294687223808 |
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| author | Lei Fang Charles Hemion David Goldblum Peter Meyer Selim Orgül Stephan Frank Josef Flammer Albert Neutzner |
| author_facet | Lei Fang Charles Hemion David Goldblum Peter Meyer Selim Orgül Stephan Frank Josef Flammer Albert Neutzner |
| author_sort | Lei Fang |
| collection | DOAJ |
| description | To study the impact of the mitochondrial ubiquitin ligase MARCH5 on mitochondrial morphology and induction of apoptosis using an in vitro model of neuronal precursor cells exposed to glaucoma-relevant stress conditions.RGC5 cells transfected with expression constructs for MARCH5, MARCH5(H43W), Dpr1(K38A) or vector control were exposed to either elevated pressure of 30 mmHg, oxidative stress caused by mitochondrial electron transport chain (ETC) inhibition, or hypoxia-reoxygenation conditions. Mitochondrial morphology of RGC5 cells was analyzed following staining of the mitochondrial marker cytochrome c and photoactivatable GFP (PAGFP) diffusion assay. Induction of apoptotic cell death in these cells was determined by analyzing the release of cytochrome c from mitochondria into the cytosol and flow cytometry.Exposure of RGC5 cells to oxidative stress conditions as well as to elevated pressure resulted in the fragmentation of the mitochondrial network in control cells as well as in cells expressing MARCH5. In cells expressing inactive MARCH5(H43W) or inactive Drp(K38A), mitochondrial fragmentation was significantly blocked and mitochondrial morphology was comparable to that of control cells under normal conditions. Exposure of RGC5 cells to elevated pressure or oxidative stress conditions induced apoptotic cell death as assessed by cytochrome c release and DNA staining, while expression of dominant-negative MARCH5(H43W) or Drp1(K38A) did significantly delay cell death.Preventing mitochondrial fragmentation through interference with the mitochondrial fission machinery protects neuronal cells from programmed cell death following exposure to stressors physiologically relevant to the pathogenesis of glaucoma. |
| first_indexed | 2024-12-23T06:56:35Z |
| format | Article |
| id | doaj.art-5a5c475f65f14d559f520229bce786c3 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-23T06:56:35Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-5a5c475f65f14d559f520229bce786c32022-12-21T17:56:18ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-01712e5263710.1371/journal.pone.0052637Inactivation of MARCH5 prevents mitochondrial fragmentation and interferes with cell death in a neuronal cell model.Lei FangCharles HemionDavid GoldblumPeter MeyerSelim OrgülStephan FrankJosef FlammerAlbert NeutznerTo study the impact of the mitochondrial ubiquitin ligase MARCH5 on mitochondrial morphology and induction of apoptosis using an in vitro model of neuronal precursor cells exposed to glaucoma-relevant stress conditions.RGC5 cells transfected with expression constructs for MARCH5, MARCH5(H43W), Dpr1(K38A) or vector control were exposed to either elevated pressure of 30 mmHg, oxidative stress caused by mitochondrial electron transport chain (ETC) inhibition, or hypoxia-reoxygenation conditions. Mitochondrial morphology of RGC5 cells was analyzed following staining of the mitochondrial marker cytochrome c and photoactivatable GFP (PAGFP) diffusion assay. Induction of apoptotic cell death in these cells was determined by analyzing the release of cytochrome c from mitochondria into the cytosol and flow cytometry.Exposure of RGC5 cells to oxidative stress conditions as well as to elevated pressure resulted in the fragmentation of the mitochondrial network in control cells as well as in cells expressing MARCH5. In cells expressing inactive MARCH5(H43W) or inactive Drp(K38A), mitochondrial fragmentation was significantly blocked and mitochondrial morphology was comparable to that of control cells under normal conditions. Exposure of RGC5 cells to elevated pressure or oxidative stress conditions induced apoptotic cell death as assessed by cytochrome c release and DNA staining, while expression of dominant-negative MARCH5(H43W) or Drp1(K38A) did significantly delay cell death.Preventing mitochondrial fragmentation through interference with the mitochondrial fission machinery protects neuronal cells from programmed cell death following exposure to stressors physiologically relevant to the pathogenesis of glaucoma.http://europepmc.org/articles/PMC3526576?pdf=render |
| spellingShingle | Lei Fang Charles Hemion David Goldblum Peter Meyer Selim Orgül Stephan Frank Josef Flammer Albert Neutzner Inactivation of MARCH5 prevents mitochondrial fragmentation and interferes with cell death in a neuronal cell model. PLoS ONE |
| title | Inactivation of MARCH5 prevents mitochondrial fragmentation and interferes with cell death in a neuronal cell model. |
| title_full | Inactivation of MARCH5 prevents mitochondrial fragmentation and interferes with cell death in a neuronal cell model. |
| title_fullStr | Inactivation of MARCH5 prevents mitochondrial fragmentation and interferes with cell death in a neuronal cell model. |
| title_full_unstemmed | Inactivation of MARCH5 prevents mitochondrial fragmentation and interferes with cell death in a neuronal cell model. |
| title_short | Inactivation of MARCH5 prevents mitochondrial fragmentation and interferes with cell death in a neuronal cell model. |
| title_sort | inactivation of march5 prevents mitochondrial fragmentation and interferes with cell death in a neuronal cell model |
| url | http://europepmc.org/articles/PMC3526576?pdf=render |
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