title: The role of Golgi reassembly and stacking protein 65 phosphorylation in H2O2-induced cell death and Golgi morphological changes.
abstract: This study aimed to investigate the effects of H2O2-induced oxidative stress on cell viability and survival, as well as changes in the distribution of Golgi apparatus and in the level of Golgi reassembly and stacking protein 65 (GRASP65). Cell viability of cultured N2a cells treated with H2O2was measured by the MTT assay. Apoptosis was measured by flow cytometry analyses. Cells labeled by indirect immunofluorescence were observed under confocal microscope to detect any Golgi morphological alterations; electron microscopy of Golgi apparatus was also done. Expression of GRASP65 and phospho-GRASP65 was examined by immunoblotting. H2O2treatment reduced the cell viability and raised the cell mortality of N2a cells in a time-dependent manner. Notable changes were only observed in the distribution and morphology of Golgi apparatus at 6 h after H2O2treatment. The expression of GRASP65 showed no significant changes at different time points; the phosphorylated GRASP65 level was significantly increased after H2O2treatment, peaked at 3 h, and finally dropped at 6 h. Taken together, GRASP65 phosphorylation may have a critical role in inducing cell death at the early stage after H2O2treatment, while its role in H2O2-induced Golgi morphological changes may be complex.
authors.affiliation: Department of Neurology, Second Hospital of Hebei Medical University, Shijiazhuang, 050000, People's Republic of China.|Department of Neurology, Second Xiangya Hospital, Central South University, Changsha, 410011, People's Republic of China.
authors.forename: Guang
authors.initials: G
authors.lastname: Ji
authors.affiliation: Department of Neurology, Second Xiangya Hospital, Central South University, Changsha, 410011, People's Republic of China.
authors.forename: Weiwei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Department of Neurology, Second Hospital of Hebei Medical University, Shijiazhuang, 050000, People's Republic of China.
authors.forename: Moyuan
authors.initials: M
authors.lastname: Quan
authors.affiliation: Department of Neurology, Second Xiangya Hospital, Central South University, Changsha, 410011, People's Republic of China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Neurology, Second Xiangya Hospital, Central South University, Changsha, 410011, People's Republic of China.
authors.forename: Hui
authors.initials: H
authors.lastname: Qu
authors.affiliation: Department of Neurology, Second Xiangya Hospital, Central South University, Changsha, 410011, People's Republic of China. huzhipinghzp@163.com.
authors.forename: Zhiping
authors.initials: Z
authors.lastname: Hu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002352:Carrier Proteins / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D006861:Hydrogen Peroxide / Q000633:toxicity*; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D018384:Oxidative Stress / Q000187:drug effects; D010766:Phosphorylation / Q000187:drug effects; D013997:Time Factors
chemical_list: D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C523888:tamalin protein, mouse; D006861:Hydrogen Peroxide
keywords: GRASP65 phosphorylation; Golgi apparatus; Golgi fragmentation; H2O2-induced oxidative stress; Neurodegenerative diseases