DOI: 10.1111/j.1460-9568.2008.06527.x. The effect of α‐synuclein knockdown on MPP+ toxicity in models of human neurons

The effect of α‐synuclein knockdown on MPP+ toxicity in models of human neurons

10.1111/j.1460-9568.2008.06527.x

Crossref journal-article

Wiley

European Journal of Neuroscience (311)

Abstract

AbstractThe protein α‐synuclein is central to the pathophysiology of Parkinson’s disease (PD) but its role in the development of neurodegeneration remains unclear. α‐Synuclein‐knockout mice develop without gross abnormality and are resistant to 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP), a mitochondrial inhibitor widely used to model parkinsonism. Here we show that differentiated human dopaminergic neuron‐like cells also have increased resistance to 1‐methyl‐4‐phenylpyridine (MPP+), the active metabolite of MPTP, when α‐synuclein is knocked down using RNA interference. In attempting to understand how this occurred we found that lowering α‐synuclein levels caused changes to intracellular vesicles, dopamine transporter (DAT) and vesicular monoamine transporter (VMAT2), each of which is known to be an important component of the early events leading to MPP+ toxicity. Knockdown of α‐synuclein reduced the availability of DAT on the neuronal surface by 50%, decreased the total number of intracellular vesicles by 37% but increased the density of VMAT2 molecules per vesicle by 2.8‐fold. However, these changes were not associated with any reduction in MPP+‐induced superoxide production, suggesting that α‐synuclein knockdown may have other downstream effects which are important. We then showed that α‐synuclein knockdown prevented MPP+‐induced activation of nitric oxide synthase (NOS). Activation of NOS is an essential step in MPTP toxicity and increasing evidence points to nitrosative stress as being important in neurodegeneration. Overall, these results show that as well as having a number of effects on cellular events upstream of mitochondrial dysfunction α‐synuclein affects pathways downstream of superoxide production, possibly involving regulation of NOS activity.

Bibliography

Fountaine, T. M., Venda, L. L., Warrick, N., Christian, H. C., Brundin, P., Channon, K. M., & WadeâMartins, R. (2008). The effect of Î±âsynuclein knockdown on MPP+ toxicity in models of human neurons. European Journal of Neuroscience, 28(12), 2459â2473. Portico.

Authors 7

Timothy M. Fountaine (first)
Lara Lourenco Venda (additional)
Nicholas Warrick (additional)
Helen C. Christian (additional)
Patrik Brundin (additional)
Keith M. Channon (additional)
Richard Wade‐Martins (additional)

References 62 Referenced 61

10.1016/S0896-6273(00)80886-7
10.1016/S0076-6879(98)96020-7
10.1016/j.niox.2006.03.008
10.1523/JNEUROSCI.22-20-08797.2002
10.1073/pnas.0406283101
10.1016/S0140-6736(04)17103-1
10.1111/j.1471-4159.1992.tb09789.x
10.1073/pnas.172514599
10.1046/j.1471-4159.2000.0742213.x
10.1002/jnr.10539
10.1016/j.neuro.2004.05.002
10.1016/j.molmed.2005.01.001
10.1002/jnr.21125
10.1126/science.290.5493.985
10.1002/jnr.20976
10.1074/jbc.M208194200
10.1016/j.pneurobio.2005.06.001
10.1016/j.neuint.2003.09.003
10.1111/j.1471-4159.2006.03907.x
10.1038/nm0996-1017
10.1002/glia.20259
10.1111/j.1460-9568.2007.05757.x
10.1021/bi972776r
10.1074/jbc.274.36.25481
10.1097/00001756-200004270-00024
10.1016/j.nbd.2005.08.018
10.1038/ng0298-106
10.1096/fsb2fj000334com
10.1093/hmg/ddg009
10.1038/70978
10.1074/jbc.M205518200
10.1006/nbdi.1997.0141
10.1006/exnr.1996.6406
10.1523/JNEUROSCI.20-09-03214.2000
10.1021/bi0487140
10.1523/JNEUROSCI.19-14-05782.1999
10.1016/j.neuint.2004.04.002
10.1523/JNEUROSCI.21-20-08053.2001
10.1016/S0169-328X(01)00257-1
10.1523/JNEUROSCI.22-08-03090.2002
10.1126/science.276.5321.2045
10.1002/(SICI)1098-2396(199809)30:1<79::AID-SYN10>3.0.CO;2-K
10.1523/JNEUROSCI.12-05-01658.1992 / J. Neurosci. / Transgenic mice with increased Cu/Zn‐superoxide dismutase activity are resistant to N‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine‐induced neurotoxicity by Przedborski S. (1992)
10.1073/pnas.93.10.4565
{'key': 'e_1_2_7_46_1', 'first-page': '135', 'article-title': 'The parkinsonian toxin MPTP: action and mechanism', 'volume': '16', 'author': 'Przedborski S.', 'year': '2000', 'journal-title': 'Restor. Neurol. Neurosci.'} / Restor. Neurol. Neurosci. / The parkinsonian toxin MPTP: action and mechanism by Przedborski S. (2000)
10.1016/S0076-6879(98)96019-0
10.1111/j.1471-4159.2004.02378.x
10.1016/S0306-4522(03)00036-8
10.1046/j.1471-4159.1995.64020936.x
10.1189/jlb.0507296
10.1002/ana.67
10.1073/pnas.94.18.9938
10.1002/j.1460-2075.1993.tb05808.x
10.1016/S0304-3940(03)00097-1
10.1096/fj.03-0152fje
10.1111/j.1460-9568.2006.05181.x
10.1111/j.1460-9568.2006.04900.x
10.1007/s00702-008-0019-6
10.1002/ana.10795
10.1093/jnen/59.1.53
10.1016/S0891-5849(03)00142-4
10.1073/pnas.0501719102

Dates

Type	When
Created	16 years, 9 months ago (Nov. 21, 2008, 6:22 a.m.)
Deposited	1 year, 10 months ago (Oct. 9, 2023, 1:59 p.m.)
Indexed	2 months, 2 weeks ago (June 17, 2025, 12:46 p.m.)
Issued	16 years, 9 months ago (Dec. 1, 2008)
Published	16 years, 9 months ago (Dec. 1, 2008)
Published Online	16 years, 8 months ago (Dec. 9, 2008)
Published Print	16 years, 9 months ago (Dec. 1, 2008)

Funders 0

None

BibTeX

@article{Fountaine_2008, title={The effect of α‐synuclein knockdown on MPP+ toxicity in models of human neurons}, volume={28}, ISSN={1460-9568}, url={http://dx.doi.org/10.1111/j.1460-9568.2008.06527.x}, DOI={10.1111/j.1460-9568.2008.06527.x}, number={12}, journal={European Journal of Neuroscience}, publisher={Wiley}, author={Fountaine, Timothy M. and Venda, Lara Lourenco and Warrick, Nicholas and Christian, Helen C. and Brundin, Patrik and Channon, Keith M. and Wade‐Martins, Richard}, year={2008}, month=dec, pages={2459–2473} }

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