PMID- 30824683 OWN - NLM STAT- MEDLINE DCOM- 20200520 LR - 20200520 IS - 2041-4889 (Electronic) VI - 10 IP - 3 DP - 2019 Mar 1 TI - Quantitative proteomic analyses of dynamic signalling events in cortical neurons undergoing excitotoxic cell death. PG - 213 LID - 10.1038/s41419-019-1445-0 [doi] LID - 213 AB - Excitotoxicity, caused by overstimulation or dysregulation of ionotropic glutamate receptors (iGluRs), is a pathological process directing neuronal death in many neurological disorders. The aberrantly stimulated iGluRs direct massive influx of calcium ions into the affected neurons, leading to changes in expression and phosphorylation of specific proteins to modulate their functions and direct their participation in the signalling pathways that induce excitotoxic neuronal death. To define these pathways, we used quantitative proteomic approaches to identify these neuronal proteins (referred to as the changed proteins) and determine how their expression and/or phosphorylation dynamically changed in association with excitotoxic cell death. Our data, available in ProteomeXchange with identifier PXD008353, identified over 100 changed proteins exhibiting significant alterations in abundance and/or phosphorylation levels at different time points (5-240 min) in neurons after glutamate overstimulation. Bioinformatic analyses predicted that many of them are components of signalling networks directing defective neuronal morphology and functions. Among them, the well-known neuronal survival regulators including mitogen-activated protein kinases Erk1/2, glycogen synthase kinase 3 (GSK3) and microtubule-associated protein (Tau), were selected for validation by biochemical approaches, which confirmed the findings of the proteomic analysis. Bioinformatic analysis predicted Protein Kinase B (Akt), c-Jun kinase (JNK), cyclin-dependent protein kinase 5 (Cdk5), MAP kinase kinase (MEK), Casein kinase 2 (CK2), Rho-activated protein kinase (Rock) and Serum/glucocorticoid-regulated kinase 1 (SGK1) as the potential upstream kinases phosphorylating some of the changed proteins. Further biochemical investigation confirmed the predictions of sustained changes of the activation states of neuronal Akt and CK2 in excitotoxicity. Thus, future investigation to define the signalling pathways directing the dynamic alterations in abundance and phosphorylation of the identified changed neuronal proteins will help elucidate the molecular mechanism of neuronal death in excitotoxicity. FAU - Hoque, Ashfaqul AU - Hoque A AUID- ORCID: 0000-0003-2384-2983 AD - Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, VIC, 3010, Australia. AD - Cell Signalling Research Laboratories, University of Melbourne, Parkville, VIC, 3010, Australia. AD - Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, VIC, 3010, Australia. AD - Metabolic Signalling Laboratory, St. Vincent's Institute for Medical Research, University of Melbourne, Fitzroy, VIC, 3065, Australia. FAU - Williamson, Nicholas A AU - Williamson NA AUID- ORCID: 0000-0002-2173-3452 AD - Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, VIC, 3010, Australia. FAU - Ameen, S Sadia AU - Ameen SS AD - Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, VIC, 3010, Australia. AD - Cell Signalling Research Laboratories, University of Melbourne, Parkville, VIC, 3010, Australia. FAU - Ciccotosto, Giuseppe D AU - Ciccotosto GD AD - Department of Pharmacology and Therapeutics, University of Melbourne, Parkville, VIC, 3010, Australia. FAU - Hossain, M Iqbal AU - Hossain MI AD - Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, VIC, 3010, Australia. AD - Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, VIC, 3010, Australia. FAU - Oakhill, Jonathan S AU - Oakhill JS AUID- ORCID: 0000-0002-9475-1440 AD - Metabolic Signalling Laboratory, St. Vincent's Institute for Medical Research, University of Melbourne, Fitzroy, VIC, 3065, Australia. AD - Mary MacKillop Institute for Health Research, Australian Catholic University, Melbourne, Victoria, 3000, Australia. FAU - Ng, Dominic C H AU - Ng DCH AD - School of Biomedical Sciences, University of Queensland, St. Lucia, QLD, Australia. FAU - Ang, Ching-Seng AU - Ang CS AD - Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, VIC, 3010, Australia. ching-seng.ang@unimelb.edu.au. FAU - Cheng, Heung-Chin AU - Cheng HC AUID- ORCID: 0000-0002-4965-7148 AD - Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, VIC, 3010, Australia. heung@unimelb.edu.au. AD - Cell Signalling Research Laboratories, University of Melbourne, Parkville, VIC, 3010, Australia. heung@unimelb.edu.au. AD - Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, VIC, 3010, Australia. heung@unimelb.edu.au. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20190301 PL - England TA - Cell Death Dis JT - Cell death & disease JID - 101524092 RN - 0 (Nerve Tissue Proteins) RN - 0 (tau Proteins) RN - 3KX376GY7L (Glutamic Acid) RN - EC 2.7.10.1 (Receptor, trkA) RN - EC 2.7.11.1 (Akt1 protein, mouse) RN - EC 2.7.11.1 (Casein Kinase II) RN - EC 2.7.11.1 (Proto-Oncogene Proteins c-akt) RN - EC 2.7.11.24 (Mapk1 protein, mouse) RN - EC 2.7.11.24 (Mitogen-Activated Protein Kinase 1) RN - EC 2.7.11.24 (Mitogen-Activated Protein Kinase 3) RN - EC 2.7.11.26 (Glycogen Synthase Kinase 3) SB - IM MH - Animals MH - Casein Kinase II/chemistry/genetics/metabolism MH - Cell Death MH - Cell Survival MH - Cells, Cultured MH - Chromatography, Liquid MH - Computational Biology MH - Glutamic Acid/metabolism/*toxicity MH - Glycogen Synthase Kinase 3/genetics/metabolism MH - Mice MH - Mitogen-Activated Protein Kinase 1/chemistry/genetics/metabolism MH - Mitogen-Activated Protein Kinase 3/chemistry/genetics/metabolism MH - Nerve Tissue Proteins/chemistry/*metabolism MH - Neurons/cytology/*drug effects/*metabolism/pathology MH - Phosphorylation MH - Proteomics MH - Proto-Oncogene Proteins c-akt/genetics/metabolism MH - Receptor, trkA/genetics/metabolism MH - Signal Transduction/*drug effects/genetics MH - Software MH - Tandem Mass Spectrometry MH - tau Proteins/chemistry/genetics/metabolism PMC - PMC6397184 COIS- The authors declare that they have no conflict of interest. EDAT- 2019/03/03 06:00 MHDA- 2020/05/21 06:00 PMCR- 2019/03/01 CRDT- 2019/03/03 06:00 PHST- 2018/10/28 00:00 [received] PHST- 2019/02/01 00:00 [accepted] PHST- 2019/01/20 00:00 [revised] PHST- 2019/03/03 06:00 [entrez] PHST- 2019/03/03 06:00 [pubmed] PHST- 2020/05/21 06:00 [medline] PHST- 2019/03/01 00:00 [pmc-release] AID - 10.1038/s41419-019-1445-0 [pii] AID - 1445 [pii] AID - 10.1038/s41419-019-1445-0 [doi] PST - epublish SO - Cell Death Dis. 2019 Mar 1;10(3):213. doi: 10.1038/s41419-019-1445-0.