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Gas1 Knockdown Increases the Neuroprotective Effect of Glial Cell-Derived Neurotrophic Factor Against Glutamate-Induced Cell Injury in Human SH-SY5Y Neuroblastoma Cells

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ABSTRACT

Growth arrest-specific 1 (Gas1) protein acts as an inhibitor of cell growth and a mediator of cell death in nervous system during development and is also re-expressed in adult neurons during excitotoxic insult. Due to its structural similarity to the glial cell-derived neurotrophic factor family receptors α (GFRα), Gas1 is likely to interfere with the neuroprotective effect of GDNF. In the present study, we investigated the expression profile of Gas1 during glutamate insults in human SH-SY5Y neuroblastoma cells as well as the influence of Gas1 inhibition on the protective effect of GDNF against glutamate-induced cell injury. Our data showed that Gas1 expression was significantly increased with the treatment of glutamate in SH-SY5Y cells. The silencing of Gas1 by small interfering RNA promoted the protective effect of GDNF against glutamate-induced cytotoxicity as well as cell apoptosis, which effect was likely mediated through activating Akt/PI3 K-dependent cell survival signaling pathway and inhibiting mitochondrial-dependent cell apoptosis signaling pathway via Bad dephosphorylation blockade. In summary, this study showed the synergistic effect of Gas1 inhibition and GDNF against glutamate-induced cell injury in human SH-SY5Y neuroblastoma cells, which information might significantly contribute to better understanding the function of Gas1 in neuronal cells and form the basis of the therapeutic development of GDNF in treating human neurodegenerative diseases in the future.

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Reference

  • Ankarcrona M, Dypbukt JM, Bonfoco E, Zhivotovsky B, Orrenius S, Lipton SA, Nicotera P (1995) Glutamate-induced neuronal death: a succession of necrosis or apoptosis depending on mitochondrial function. Neuron 15:961–973

    Article  CAS  PubMed  Google Scholar 

  • Beal MF (1992) Mechanisms of excitotoxicity in neurologic diseases. FASEB J 6:3338–3344

    CAS  PubMed  Google Scholar 

  • Bonde C, Kristensen BW, Blaabjerg M, Johansen TE, Zimmer J, Meyer M (2000) GDNF and neublastin protect against NMDA-induced excitotoxicity in hippocampal slice cultures. NeuroReport 11:4069–4073

    Article  CAS  PubMed  Google Scholar 

  • Cabrera JR, Sanchez-Pulido L, Rojas AM, Valencia A, Mañes S, Naranjo JR, Mellström B (2006) Gas1 is related to the glial cell-derived neurotrophic factor family receptors α and regulates Ret signaling. J Biol Chem 281:14330–14339

    Article  CAS  PubMed  Google Scholar 

  • Chalak LF, Rouse DJ (2011) Neuroprotective approaches: before and after delivery. Clin Perinatol 38:455–470

    Article  PubMed  Google Scholar 

  • Choi-Lundberg DL, Lin Q, Chang Y-N, Chiang YL, Hay CM, Mohajeri H, Davidson BL, Bohn MC (1997) Dopaminergic neurons protected from degeneration by GDNF gene therapy. Science 275:838–841

    Article  CAS  PubMed  Google Scholar 

  • Coyle JT, Puttfarcken P (1993) Oxidative stress, glutamate, and neurodegenerative disorders. Science 262:689–695

    Article  CAS  PubMed  Google Scholar 

  • Datta SR, Dudek H, Tao X, Masters S, Fu H, Gotoh Y, Greenberg ME (1997) Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell 91:231–241

    Article  CAS  PubMed  Google Scholar 

  • Del Sal G, Ruaro ME, Philipson L, Schneider C (1992) The growth arrest-specific gene, gas1, is involved in growth suppression. Cell 70:595–607

    Article  PubMed  Google Scholar 

  • Edgar D, Barde Y-A (1983) Neuronal growth factors. Trends Neurosci 6:260–262

    Article  CAS  Google Scholar 

  • Evdokiou A, Cowled PA (1998) Tumor-suppressive activity of the growth arrest-specific gene GAS1 in human tumor cell lines. Int J Cancer 75:568–577

    Article  CAS  PubMed  Google Scholar 

  • Ghribi O, Herman MM, Forbes MS, DeWitt DA, Savory J (2001) GDNF protects against aluminum-induced apoptosis in rabbits by upregulating Bcl-2 and Bcl-XL and inhibiting mitochondrial Bax translocation. Neurobiology of Disease 8:764–773

    Article  CAS  PubMed  Google Scholar 

  • Gibson GE, Starkov A, Blass JP, Ratan RR, Beal MF (2010) Cause and consequence: mitochondrial dysfunction initiates and propagates neuronal dysfunction, neuronal death and behavioral abnormalities in age-associated neurodegenerative diseases. Biochim Biophys Acta 1802:122–134

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gottlieb TM, Leal J, Seger R, Taya Y, Oren M (2002) Cross-talk between Akt, p53 and Mdm2: possible implications for the regulation of apoptosis. Oncogene 21:1299–1303

    Article  CAS  PubMed  Google Scholar 

  • Gratacòs E, Pérez-Navarro E, Tolosa E, Arenas E, Alberch J (2001) Neuroprotection of striatal neurons against kainate excitotoxicity by neurotrophins and GDNF family members. J Neurochem 78:1287–1296

    Article  PubMed  Google Scholar 

  • Greenamyre JT, Porter R (1994) Anatomy and physiology of glutamate in the CNS. Neurology 44:S7–S13

    CAS  PubMed  Google Scholar 

  • Hansford LM, Marshall GM (2005) Glial cell line-derived neurotrophic factor (GDNF) family ligands reduce the sensitivity of neuroblastoma cells to pharmacologically induced cell death, growth arrest and differentiation. Neurosci Lett 389:77–82

    Article  CAS  PubMed  Google Scholar 

  • Kruger NJ (1994) The Bradford method for protein quantitation. Methods Mol Biol 32:9–15

    CAS  PubMed  Google Scholar 

  • Lang AE, Gill S, Patel NK, Lozano A, Nutt JG, Penn R, Brooks DJ, Hotton G, Moro E, Heywood P (2006) Randomized controlled trial of intraputamenal glial cell line–derived neurotrophic factor infusion in Parkinson disease. Ann Neurol 59:459–466

    Article  CAS  PubMed  Google Scholar 

  • Lemasters JJ, Qian T, Bradham CA, Brenner DA, Cascio WE, Trost LC, Nishimura Y, Nieminen A-L, Herman B (1999) Mitochondrial dysfunction in the pathogenesis of necrotic and apoptotic cell death. J Bioenerg Biomembr 31:305–319

    Article  CAS  PubMed  Google Scholar 

  • Lin LF, Doherty DH, Lile JD, Bektesh S, Collins F (1993) GDNF: a glial cell line-derived neurotrophic factor for midbrain dopaminergic neurons. Science 260:1130–1132

    Article  CAS  PubMed  Google Scholar 

  • López-Ramírez MA, Domínguez-Monzón G, Vergara P, Segovia J (2008) Gas1 reduces Ret tyrosine 1062 phosphorylation and alters GDNF-mediated intracellular signaling. Int J Dev Neurosci 26:497–503

    Article  PubMed  Google Scholar 

  • Martinelli DC, Fan C-M (2007) The role of Gas1 in embryonic development and its implications for human disease. Cell Cycle 6:2650–2655

    Article  CAS  PubMed  Google Scholar 

  • Maruyama W, Nitta A, Shamoto-Nagai M, Hirata Y, Akao Y, Yodim M, Furukawa S, Nabeshima T, Naoi M (2004) N-Propargyl-1 (R)-aminoindan, rasagiline, increases glial cell line-derived neurotrophic factor (GDNF) in neuroblastoma SH-SY5Y cells through activation of NF-κB transcription factor. Neurochem Int 44:393–400

    Article  CAS  PubMed  Google Scholar 

  • Mellstrom B, Cena V, Lamas M, Perales C, Gonzalez C, Naranjo JR (2002) Gas1 is induced during and participates in excitotoxic neuronal death. Mol Cell Neurosci 19:417–429

    Article  PubMed  Google Scholar 

  • Nutt J, Burchiel K, Comella C, Jankovic J, Lang A, Laws E, Lozano A, Penn R, Simpson R, Stacy M (2003) Randomized, double-blind trial of glial cell line-derived neurotrophic factor (GDNF) in PD. Neurology 60:69–73

    Article  CAS  PubMed  Google Scholar 

  • Patel NK, Bunnage M, Plaha P, Svendsen CN, Heywood P, Gill SS (2005) Intraputamenal infusion of glial cell line–derived neurotrophic factor in PD: a two-year outcome study. Ann Neurol 57:298–302

    Article  CAS  PubMed  Google Scholar 

  • Pizza V, Agresta A, D’Acunto CW, Festa M, Capasso A (2011) Neuroinflamm-aging and neurodegenerative diseases: an overview. CNS Neurol Disord Drug Targets 10:621–634

    Article  CAS  PubMed  Google Scholar 

  • Saarma M (2000) GDNF—a stranger in the TGF-β superfamily? Eur J Biochem 267:6968–6971

    Article  CAS  PubMed  Google Scholar 

  • Saxena S, Caroni P (2011) Selective neuronal vulnerability in neurodegenerative diseases: from stressor thresholds to degeneration. Neuron 71:35–48

    Article  CAS  PubMed  Google Scholar 

  • Schueler-Furman O, Glick E, Segovia J, Linial M (2006) Is GAS1 a co-receptor for the GDNF family of ligands? Trends Pharmacol Sci 27:72–77

    Article  CAS  PubMed  Google Scholar 

  • Sherer TB, Fiske BK, Svendsen CN, Lang AE, Langston JW (2006) Crossroads in GDNF therapy for Parkinson’s disease. Mov Disord 21:136–141

    Article  PubMed  Google Scholar 

  • Treanor JJ, Goodman L, de Sauvage F, Stone DM, Poulsen KT, Beck CD, Gray C, Armanini MP, Pollock RA, Hefti F, Phillips HS, Goddard A, Moore MW, Buj-Bello A, Davies AM, Asai N, Takahashi M, Vandlen R, Henderson CE, Rosenthal A (1996) Characterization of a multicomponent receptor for GDNF. Nature 382:80–83

    Article  CAS  PubMed  Google Scholar 

  • Ueda K, Fukui Y, Kageyama H (1994) Amyloid β protein-induced neuronal cell death: neurotoxic properties of aggregated amyloid β protein. Brain Res 639:240–244

    Article  CAS  PubMed  Google Scholar 

  • van Meerloo J, Kaspers GJ, Cloos J (2011) Cell sensitivity assays: the MTT assay. Methods Mol Biol 731:237–245

    Article  PubMed  Google Scholar 

  • Wang Y, Qin Z-H (2010) Molecular and cellular mechanisms of excitotoxic neuronal death. Apoptosis 15:1382–1402

    Article  CAS  PubMed  Google Scholar 

  • Yan Q, Matheson C, Lopez OT (1995) In vivo neurotrophic effects of GDNF on neonatal and adult facial motor neurons. Nature 373:341–344

    Article  CAS  PubMed  Google Scholar 

  • Yu F, Sugawara T, Maier CM, Hsieh LB, Chan PH (2005) Akt/Bad signaling and motor neuron survival after spinal cord injury. Neurobiol Dis 20:491–499

    Article  CAS  PubMed  Google Scholar 

  • Yuan J, Lipinski M, Degterev A (2003) Diversity in the mechanisms of neuronal cell death. Neuron 40:401–413

    Article  CAS  PubMed  Google Scholar 

  • Zarco N, González-Ramírez R, González RO, Segovia J (2012) GAS1 induces cell death through an intrinsic apoptotic pathway. Apoptosis 17:627–635

    Article  CAS  PubMed  Google Scholar 

  • Zarco N, Bautista E, Cuellar M, Vergara P, Flores-Rodriguez P, Aguilar-Roblero R, Segovia J (2013) Growth arrest specific 1 (GAS1) is abundantly expressed in the adult mouse central nervous system. J Histochem Cytochem 61:731–748

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhu X, Wang K, Zhang K, Zhu L, Zhou F (2014) Ziyuglycoside II induces cell cycle arrest and apoptosis through activation of ROS/JNK pathway in human breast cancer cells. Toxicol Lett 227:65–73

    Article  CAS  PubMed  Google Scholar 

  • Zundel W, Giaccia A (1998) Inhibition of the anti-apoptotic PI (3) K/Akt/Bad pathway by stress. Genes Dev 12:1941–1946

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the National Natural Science Foundation (81300787), the Natural Science Foundation of Jiangsu Province (BK2011168, BK2012105, BK20141103), and the Major Project of Wuxi Municipal Health Bureau (ZS201401).

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Authors and Affiliations

  1. Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Qianrong Road 20, Wuxi, 214063, Jiangsu, China

    Ke Wang, Xue Zhu & Kai Zhang

  2. Faculty of Pharmacy, University of Sydney, Room 104 Medical Foundation Building K25, Sydney, NSW, 2006, Australia

    Fanfan Zhou

  3. Save Sight Institute, University of Sydney, Sydney, NSW, 2000, Australia

    Ling Zhu

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  1. Ke Wang
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  2. Xue Zhu
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  3. Kai Zhang
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  5. Ling Zhu
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Correspondence to Ke Wang or Fanfan Zhou.

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Wang, K., Zhu, X., Zhang, K. et al. Gas1 Knockdown Increases the Neuroprotective Effect of Glial Cell-Derived Neurotrophic Factor Against Glutamate-Induced Cell Injury in Human SH-SY5Y Neuroblastoma Cells. Cell Mol Neurobiol 36, 603–611 (2016). https://doi.org/10.1007/s10571-015-0241-3

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