title: Roles of Neuroglobin Binding to Mitochondrial Complex III Subunit Cytochrome c1 in Oxygen-Glucose Deprivation-Induced Neurotoxicity in Primary Neurons.
abstract: Neuroglobin (Ngb) is a tissue globin specifically expressed in brain neurons. Recent studies by our laboratory and others have demonstrated that Ngb is protective against stroke and related neurological disorders, but the mechanisms remain poorly understood. We previously identified cytochrome c1 (Cyc1) as an Ngb-interacting molecule by yeast two-hybrid screening. Cyc1 is a subunit of mitochondria complex III, which is a component of mitochondrial respiratory chain and a major source of reactive oxygen species (ROS) production under both physiological and pathological conditions. In this study, we for the first time defined Ngb-Cyc1 binding, and investigated its roles in oxygen-glucose deprivation (OGD)/reoxygenation-induced neurotoxicity and ROS production in primary neurons. Immunocytochemistry and co-immunoprecipitation validated Ngb-Cyc1 binding, which was significantly increased by OGD and Ngb overexpression. We found 4 h OGD with/without 4 h reoxygenation significantly increased complex III activity, but this activity elevation was significantly attenuated in three groups of neurons: Ngb overexpression, specific complex III inhibitor stigmatellin, or stigmatellin plus Ngb overexpression, whereas there was no significant differences between these three groups, suggesting Ngb-Cyc1 binding may function in suppressing OGD-mediated complex III activity elevation. Importantly, these three groups of neurons also showed significant decreases in OGD-induced superoxide anion generation and neurotoxicity. These results suggest that Ngb can bind to mitochondrial complex III subunit Cyc1, leading to suppression of OGD-mediated complex III activity and subsequent ROS production elevation, and eventually reduction of OGD-induced neurotoxicity. This molecular signaling cascade may be at least part of the mechanisms of Ngb neuroprotection against OGD-induced neurotoxicity.
authors.affiliation: Department of Neurosurgery, The First Affiliated Hospital, College of Pharmacy, Wenzhou Medical University, Wenzhou, Zhejiang, 325000, China. yuzywzmu@126.com.|Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School, 149 13th Street, Room 2411A, Charlestown, MA, 02129, USA. yuzywzmu@126.com.
authors.forename: Zhanyang
authors.initials: Z
authors.lastname: Yu
authors.affiliation: Department of Neurosurgery, The First Affiliated Hospital, College of Pharmacy, Wenzhou Medical University, Wenzhou, Zhejiang, 325000, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Protein Biochemistry and Developmental Biology of State Education Ministry, College of Life Sciences, Hunan Normal University, Changsha, Hunan, 410081, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Liu
authors.affiliation: Key Laboratory of Protein Biochemistry and Developmental Biology of State Education Ministry, College of Life Sciences, Hunan Normal University, Changsha, Hunan, 410081, China.|Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School, 149 13th Street, Room 2411A, Charlestown, MA, 02129, USA.
authors.forename: Jing
authors.initials: J
authors.lastname: Yuan
authors.affiliation: Department of Neurosurgery, The First Affiliated Hospital, College of Pharmacy, Wenzhou Medical University, Wenzhou, Zhejiang, 325000, China.
authors.forename: Li
authors.initials: L
authors.lastname: Lin
authors.affiliation: Department of Neurosurgery, The First Affiliated Hospital, College of Pharmacy, Wenzhou Medical University, Wenzhou, Zhejiang, 325000, China.
authors.forename: Qichuan
authors.initials: Q
authors.lastname: Zhuge
authors.affiliation: Department of Neurosurgery, The First Affiliated Hospital, College of Pharmacy, Wenzhou Medical University, Wenzhou, Zhejiang, 325000, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Xiao
authors.affiliation: Neuroprotection Research Laboratory, Departments of Neurology and Radiology, Massachusetts General Hospital, and Program in Neuroscience, Harvard Medical School, 149 13th Street, Room 2411A, Charlestown, MA, 02129, USA. wangxi@helix.mgh.harvard.edu.
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Wang
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003575:Cytochromes c1 / Q000378:metabolism*; D014450:Electron Transport Complex III / Q000378:metabolism; D005260:Female; D005914:Globins / Q000378:metabolism*; D005947:Glucose / Q000172:deficiency*; D008810:Mice, Inbred C57BL; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D000078787:Neuroglobin; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D020258:Neurotoxicity Syndromes / Q000378:metabolism* / Q000473:pathology*; D010100:Oxygen / Q000378:metabolism*; D011090:Polyenes / Q000494:pharmacology; D011485:Protein Binding / Q000187:drug effects; D021122:Protein Subunits / Q000378:metabolism*; D015203:Reproducibility of Results; D013481:Superoxides / Q000378:metabolism
chemical_list: D009419:Nerve Tissue Proteins; D000078787:Neuroglobin; D011090:Polyenes; D021122:Protein Subunits; D013481:Superoxides; D005914:Globins; D003575:Cytochromes c1; C041573:stigmatellin; D014450:Electron Transport Complex III; D005947:Glucose; D010100:Oxygen
keywords: Complex III; Cytochrome c1; Neuroglobin; Oxygen-glucose deprivation; Primary neurons; Superoxide anion