Neuroglobin overexpression inhibits oxygen–glucose deprivation-induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neurons
Neuroglobin (Ngb) is an endogenous neuroprotective molecule against hypoxic/ischemic brain injury, but the underlying mechanisms remain largely undefined. Our recent study revealed that Ngb can bind to voltage-dependent anion channel (VDAC), a regulator of mitochondria permeability transition (MPT)....
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2013-08-01
|
| Series: | Neurobiology of Disease |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0969996113001290 |
| _version_ | 1818869980173697024 |
|---|---|
| author | Zhanyang Yu Ning Liu Yadan Li Jianfeng Xu Xiaoying Wang |
| author_facet | Zhanyang Yu Ning Liu Yadan Li Jianfeng Xu Xiaoying Wang |
| author_sort | Zhanyang Yu |
| collection | DOAJ |
| description | Neuroglobin (Ngb) is an endogenous neuroprotective molecule against hypoxic/ischemic brain injury, but the underlying mechanisms remain largely undefined. Our recent study revealed that Ngb can bind to voltage-dependent anion channel (VDAC), a regulator of mitochondria permeability transition (MPT). In this study we examined the role of Ngb in MPT pore (mPTP) opening following oxygen–glucose deprivation (OGD) in primary cultured mouse cortical neurons. Co-immunoprecipitation (Co-IP) and immunocytochemistry showed that the binding between Ngb and VDAC was increased after OGD compared to normoxia, indicating the OGD-enhanced Ngb–VDAC interaction. Ngb overexpression protected primary mouse cortical neurons from OGD-induced neuronal death, to an extent comparable to mPTP opening inhibitor, cyclosporine A (CsA) pretreatment. We further measured the role of Ngb in OGD-induced mPTP opening using Ngb overexpression and knockdown approaches in primary cultured neurons, and recombinant Ngb exposure to isolated mitochondria. Same as CsA pretreatment, Ngb overexpression significantly reduced OGD-induced mPTP opening markers including mitochondria swelling, mitochondrial NAD+ release, and cytochrome c (Cyt c) release in primary cultured neurons. Recombinant Ngb incubation significantly reduced OGD-induced NAD+ release and Cyt c release from isolated mitochondria. In contrast, Ngb knockdown significantly increased OGD-induced neuron death, and increased OGD-induced mitochondrial NAD+ release and Cyt c release as well, and these outcomes could be rescued by CsA pretreatment. In summary, our results demonstrated that Ngb overexpression can inhibit OGD-induced mPTP opening in primary cultured mouse cortical neurons, which may be one of the molecular mechanisms of Ngb's neuroprotection. |
| first_indexed | 2024-12-19T11:59:44Z |
| format | Article |
| id | doaj.art-9097453e84d944298b71bd06b6e40d7e |
| institution | Directory Open Access Journal |
| issn | 1095-953X |
| language | English |
| last_indexed | 2024-12-19T11:59:44Z |
| publishDate | 2013-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Neurobiology of Disease |
| spelling | doaj.art-9097453e84d944298b71bd06b6e40d7e2022-12-21T20:22:32ZengElsevierNeurobiology of Disease1095-953X2013-08-015695103Neuroglobin overexpression inhibits oxygen–glucose deprivation-induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neuronsZhanyang Yu0Ning Liu1Yadan Li2Jianfeng Xu3Xiaoying Wang4Correspondence to: Z. Yu, Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, Harvard Medical School, 149 13th Street, Room 2401, Charlestown, MA 02129, USA. Fax: +1 617 726 7830.; Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, Charlestown, MA 02129, USA; Program in Neuroscience, Harvard Medical School, Charlestown, MA 02129, USANeuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, Charlestown, MA 02129, USA; Program in Neuroscience, Harvard Medical School, Charlestown, MA 02129, USANeuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, Charlestown, MA 02129, USA; Program in Neuroscience, Harvard Medical School, Charlestown, MA 02129, USANeuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, Charlestown, MA 02129, USA; Program in Neuroscience, Harvard Medical School, Charlestown, MA 02129, USACorrespondence to: X. Wang, Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, Harvard Medical School, 149 13th Street, Room 2411A, Charlestown, MA 02129, USA. Fax: +1 617 726 7830.; Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, Charlestown, MA 02129, USA; Program in Neuroscience, Harvard Medical School, Charlestown, MA 02129, USANeuroglobin (Ngb) is an endogenous neuroprotective molecule against hypoxic/ischemic brain injury, but the underlying mechanisms remain largely undefined. Our recent study revealed that Ngb can bind to voltage-dependent anion channel (VDAC), a regulator of mitochondria permeability transition (MPT). In this study we examined the role of Ngb in MPT pore (mPTP) opening following oxygen–glucose deprivation (OGD) in primary cultured mouse cortical neurons. Co-immunoprecipitation (Co-IP) and immunocytochemistry showed that the binding between Ngb and VDAC was increased after OGD compared to normoxia, indicating the OGD-enhanced Ngb–VDAC interaction. Ngb overexpression protected primary mouse cortical neurons from OGD-induced neuronal death, to an extent comparable to mPTP opening inhibitor, cyclosporine A (CsA) pretreatment. We further measured the role of Ngb in OGD-induced mPTP opening using Ngb overexpression and knockdown approaches in primary cultured neurons, and recombinant Ngb exposure to isolated mitochondria. Same as CsA pretreatment, Ngb overexpression significantly reduced OGD-induced mPTP opening markers including mitochondria swelling, mitochondrial NAD+ release, and cytochrome c (Cyt c) release in primary cultured neurons. Recombinant Ngb incubation significantly reduced OGD-induced NAD+ release and Cyt c release from isolated mitochondria. In contrast, Ngb knockdown significantly increased OGD-induced neuron death, and increased OGD-induced mitochondrial NAD+ release and Cyt c release as well, and these outcomes could be rescued by CsA pretreatment. In summary, our results demonstrated that Ngb overexpression can inhibit OGD-induced mPTP opening in primary cultured mouse cortical neurons, which may be one of the molecular mechanisms of Ngb's neuroprotection.http://www.sciencedirect.com/science/article/pii/S0969996113001290NeuroglobinOxygen/glucose deprivationMitochondria permeability transition pore (mPTP)NAD+ releaseMitochondria swellingCytochrome c |
| spellingShingle | Zhanyang Yu Ning Liu Yadan Li Jianfeng Xu Xiaoying Wang Neuroglobin overexpression inhibits oxygen–glucose deprivation-induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neurons Neurobiology of Disease Neuroglobin Oxygen/glucose deprivation Mitochondria permeability transition pore (mPTP) NAD+ release Mitochondria swelling Cytochrome c |
| title | Neuroglobin overexpression inhibits oxygen–glucose deprivation-induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neurons |
| title_full | Neuroglobin overexpression inhibits oxygen–glucose deprivation-induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neurons |
| title_fullStr | Neuroglobin overexpression inhibits oxygen–glucose deprivation-induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neurons |
| title_full_unstemmed | Neuroglobin overexpression inhibits oxygen–glucose deprivation-induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neurons |
| title_short | Neuroglobin overexpression inhibits oxygen–glucose deprivation-induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neurons |
| title_sort | neuroglobin overexpression inhibits oxygen glucose deprivation induced mitochondrial permeability transition pore opening in primary cultured mouse cortical neurons |
| topic | Neuroglobin Oxygen/glucose deprivation Mitochondria permeability transition pore (mPTP) NAD+ release Mitochondria swelling Cytochrome c |
| url | http://www.sciencedirect.com/science/article/pii/S0969996113001290 |
| work_keys_str_mv | AT zhanyangyu neuroglobinoverexpressioninhibitsoxygenglucosedeprivationinducedmitochondrialpermeabilitytransitionporeopeninginprimaryculturedmousecorticalneurons AT ningliu neuroglobinoverexpressioninhibitsoxygenglucosedeprivationinducedmitochondrialpermeabilitytransitionporeopeninginprimaryculturedmousecorticalneurons AT yadanli neuroglobinoverexpressioninhibitsoxygenglucosedeprivationinducedmitochondrialpermeabilitytransitionporeopeninginprimaryculturedmousecorticalneurons AT jianfengxu neuroglobinoverexpressioninhibitsoxygenglucosedeprivationinducedmitochondrialpermeabilitytransitionporeopeninginprimaryculturedmousecorticalneurons AT xiaoyingwang neuroglobinoverexpressioninhibitsoxygenglucosedeprivationinducedmitochondrialpermeabilitytransitionporeopeninginprimaryculturedmousecorticalneurons |