title: Proteomic analysis of PSD-93 knockout mice following the induction of ischemic cerebral injury.
abstract: Postsynaptic density protein-93 (PSD-93) is enriched in the postsynaptic density and is involved in N-methyl-d-aspartate receptor (NMDAR) triggered neurotoxicity through PSD-93/NMDAR/nNOS signaling pathway. In the present study, we found that PSD-93 deficiency reduced infarcted volume and neurological deficits induced by transient middle cerebral artery occlusion (tMCAO) in the mice. To identify novel targets of PSD-93 related neurotoxicity, we applied isobaric tags for relative and absolute quantitative (iTRAQ) labeling and combined this labeling with on-line two-dimensional LC/MS/MS technology to elucidate the changes in protein expression in PSD-93 knockout mice following tMCAO. The proteomic data set consisted of 1892 proteins. Compared to control group, differences in expression levels in ischemic group >1.5-fold and <0.66-fold were considered as differential expression. A total of 104 unique proteins with differential abundance levels were identified, among which 17 proteins were selected for further validation. Gene ontology analysis using UniProt database revealed that these differentially expressed proteins are involved in diverse function such as synaptic transmission, neuronal neurotransmitter and ion transport, modification of organelle membrane components. Moreover, network analysis revealed that the interacting proteins were involved in the transport of synaptic vesicles, the integrity of synaptic membranes and the activation of the ionotropic glutamate receptors NMDAR1 and NMDAR2B. Finally, RT-PCR and Western blot analysis showed that SynGAP, syntaxin-1A, protein kinase C β, and voltage-dependent L-type calcium channels were inhibited by ischemia-reperfusion. Identification of these proteins provides valuable clues to elucidate the mechanisms underlying the actions of PSD-93 in ischemia-reperfusion induced neurotoxicity.
authors.affiliation: Department of Neurology, Drum Tower Hospital of Nanjing Medical University, Nanjing, 210029 Jiangsu, China; Department of Neurology, Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, 210029 Jiangsu, China; Jiangsu Key Laboratory for Molecular Medicine, Nanjing, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Rong
authors.affiliation: Department of Neurosurgery, Xuzhou First People's Hospital, Xuzhou, 221000 Jiangsu, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Yang
authors.affiliation: Department of Neurology, Second Affiliated Hospital of Xuzhou Medical College, Xuzhou, 221006 Jiangsu, China.
authors.forename: Liangqun
authors.initials: L
authors.lastname: Rong
authors.affiliation: Department of Neurology, Second Affiliated Hospital of Xuzhou Medical College, Xuzhou, 221006 Jiangsu, China.
authors.forename: Xiue
authors.initials: X
authors.lastname: Wei
authors.affiliation: Department of Neurology, Affiliated Hospital of Xuzhou Medical College, Xuzhou, 221000 Jiangsu, China.
authors.forename: Qingjie
authors.initials: Q
authors.lastname: Li
authors.affiliation: Department of Pathogenic Biology and Immunology, Lab of Infection and Immunity, Xuzhou Medical College, Xuzhou, 221004 Jiangsu, China.
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Neurology, Second Affiliated Hospital of Xuzhou Medical College, Xuzhou, 221006 Jiangsu, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Gao
authors.affiliation: Department of Neurology, Drum Tower Hospital of Nanjing Medical University, Nanjing, 210029 Jiangsu, China; Department of Neurology, Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, 210029 Jiangsu, China; Jiangsu Key Laboratory for Molecular Medicine, Nanjing, China. Electronic address: xuyun20042001@aliyun.com.
authors.forename: Yun
authors.initials: Y
authors.lastname: Xu
authors.affiliation: Department of Neurology, Second Affiliated Hospital of Xuzhou Medical College, Xuzhou, 221006 Jiangsu, China. Electronic address: zhangqingxiu@163.com.
authors.forename: Qingxiu
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D000818:Animals; D015220:Calcium Channels / Q000378:metabolism; D004195:Disease Models, Animal; D005786:Gene Expression Regulation / Q000235:genetics*; D063990:Gene Ontology; D051528:Guanylate Kinases / Q000172:deficiency* / Q000235:genetics; D020244:Infarction, Middle Cerebral Artery / Q000150:complications / Q000378:metabolism* / Q000473:pathology; D008565:Membrane Proteins / Q000172:deficiency* / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009422:Nervous System Diseases / Q000209:etiology / Q000235:genetics; D060066:Protein Interaction Maps / Q000235:genetics; D011493:Protein Kinase C / Q000378:metabolism; D040901:Proteomics; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism; D013223:Statistics as Topic; D050827:Syntaxin 1 / Q000378:metabolism; D020703:ras GTPase-Activating Proteins / Q000378:metabolism
chemical_list: D015220:Calcium Channels; D051528:Guanylate Kinases; D008565:Membrane Proteins; D011493:Protein Kinase C; D016194:Receptors, N-Methyl-D-Aspartate; D050827:Syntaxin 1; D020703:ras GTPase-Activating Proteins; C121001:NR2B NMDA receptor; C484512:Syngap1 protein, mouse; C501445:Dlg2 protein, mouse
keywords: Excitotoxicity; Ischemic stroke; PSD-93; Proteomic analysis