Overexpression of human α-synuclein causes dopamine neuron death in primary human mesencephalic culture

Zhou, Wenbo; Schaack, Jerome; Zawada, W. Michael; Freed, Curt R.

doi:10.1016/s0006-8993(01)03292-9

Cited by 133 publications

(89 citation statements)

References 41 publications

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“…5, B and C), we chose not to introduce the WT protein. However, we cannot exclude the possibility that overexpression of the WT protein would have resulted in DA-related toxicity, as suggested by a recent study (44).…”

Section: Discussionmentioning

confidence: 81%

“…HEK293) (54) or rat mesencephalic cells (53). Studies by Zhou and colleagues (44,59) demonstrate that the pathogenicity of A53TSYN is not only dependent on the cell type used but on the species in which the mutant protein is expressed. For instance, although adenovirus-mediated overexpression of A53T mutant human ␣-synuclein in rat primary mesencephalic cultures led to a Ն60% decrease in TH cell viability compared with cultures transduced with GFP (59), human primary mesencephalic cultures transduced with the same vector displayed only a Յ20% loss of TH-positive cells (44).…”

Section: Discussionmentioning

confidence: 99%

“…Therefore we chose not to overexpress WT human ␣-synuclein in control cultures, even though the possibility that overexpression of the WT protein itself leads to pathogenicity cannot be excluded. Indeed, a recent study showed that adenoviral-induced overexpression of WT human ␣-synuclein in primary human mesencephalic cultures increased the rate of apoptosis in transduced DA neurons by 2-fold (44). In the present study, we used an m.o.i.…”

Section: Fig 2 Differentiated Mesc210 Cultures Display Markers Of mentioning

confidence: 99%

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Effect of Mutant α-Synuclein on Dopamine Homeostasis in a New Human Mesencephalic Cell Line

Lotharius¹,

Barg²,

Wiekop³

et al. 2002

Journal of Biological Chemistry

326

266

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Mutations in ␣-synuclein have been linked to rare, autosomal dominant forms of Parkinson's disease. Despite its ubiquitous expression, mutant ␣-synuclein primarily leads to the loss of dopamine-producing neurons in the substantia nigra. ␣-Synuclein is a presynaptic nerve terminal protein of unknown function, although several studies suggest it is important for synaptic plasticity and maintenance. The present study utilized a new human mesencephalic cell line, MESC2.10, to study the effect of A53T mutant ␣-synuclein on dopamine homeostasis. In addition to expressing markers of mature dopamine neurons, differentiated MESC2.10 cells are electrically active, produce dopamine, and express wildtype human ␣-synuclein. Lentivirus-induced overexpression of A53T mutant ␣-synuclein in differentiated MESC2.10 cells resulted in down-regulation of the vesicular dopamine transporter (VMAT2), decreased potassium-induced and increased amphetamine-induced dopamine release, enhanced cytoplasmic dopamine immunofluorescence, and increased intracellular levels of superoxide. These results suggest that mutant ␣-synuclein leads to an impairment in vesicular dopamine storage and consequent accumulation of dopamine in the cytosol, a pathogenic mechanism that underlies the toxicity of the psychostimulant amphetamine and the parkinsonian neurotoxin 1-methyl-4-phenylpyridinium. Interestingly, cells expressing A53T mutant ␣-synuclein were resistant to amphetamine-induced toxicity. Because extravesicular, cytoplasmic dopamine can be easily oxidized into reactive oxygen species and other toxic metabolites, mutations in ␣-synuclein might lead to Parkinson's disease by triggering protracted, low grade dopamine toxicity resulting in terminal degeneration and ultimately cell death.

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Section: Discussionmentioning

confidence: 81%

Section: Discussionmentioning

confidence: 99%

Section: Fig 2 Differentiated Mesc210 Cultures Display Markers Of mentioning

confidence: 99%

See 1 more Smart Citation

Effect of Mutant α-Synuclein on Dopamine Homeostasis in a New Human Mesencephalic Cell Line

Lotharius¹,

Barg²,

Wiekop³

et al. 2002

Journal of Biological Chemistry

326

266

View full text Add to dashboard Cite

show abstract

“…Besides these known p53 target genes, c-Myc repressed other genes, whose expression was augmented in K562 expressing wild-type conformation p53 (32 genes included in Table 2). Among these, there are several genes previously associated with cell death induction, as a-synuclein (Iwata et al, 2001;Zhou et al, 2002) and renin (Pierzchalski et al, 1997).…”

Section: How Does C-myc Impair the P53-mediated Apoptosis In K562 Cells?mentioning

confidence: 99%

Inhibitory effect of c-Myc on p53-induced apoptosis in leukemia cells. Microarray analysis reveals defective induction of p53 target genes and upregulation of chaperone genes

et al. 2005

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We have previously demonstrated that c-Myc impairs p53-mediated apoptosis in K562 human leukemia cells, which lack ARF. To investigate the mechanisms by which c-Myc protects from p53-mediated apoptosis, we used K562 cells that conditionally express c-Myc and harbor a temperature-sensitive allele of p53. Gene expression profiles of cells expressing wild-type conformation p53 in the presence of either uninduced or induced c-Myc were analysed by cDNA microarrays. The results show that multiple p53 target genes are downregulated when c-Myc is present, including p21 WAF1 , MDM2, PERP, NOXA, GADD45, DDB2, PIR121 and p53R2. Also, a number of genes that are upregulated by c-Myc in cells expressing wild-type conformation p53 encode chaperones related to cell death protection as HSP105, HSP90 and HSP27. Both downregulation of p53 target genes and upregulation of chaperones could explain the inhibition of apoptosis observed in K562 cells with ectopic c-Myc. Myc-mediated impairment of p53 transactivation was not restricted to K562 cells, but it was reproduced in a panel of human cancer cell lines derived from different tissues. Our data suggest that elevated levels of Myc counteract p53 activity in human tumor cells that lack ARF. This mechanism could contribute to explain the c-Myc deregulation frequently found in cancer.

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“…Mutant α-syn or overexpression of wildtype α-syn induces cell death in dopaminergic cell lines and in primary dopaminergic cultures (Oluwatosin-Chigbu et al, 2003;Xu et al, 2002;Zhou et al, 2000;Zhou et al, 2002). Transgenic mice expressing mutant or wildtype α-syn show motor deficits, alterations in dopamine levels, and α-syn-positive inclusions (Hashimoto et al, 2003;Maries et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

Transcriptional dysregulation in a transgenic model of Parkinson disease

Yacoubian

Cantuti-Castelvetri

Bouzou

et al. 2008

Neurobiology of Disease

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Alpha-synuclein has been implicated in Parkinson's disease, yet the mechanism by which alphasynuclein causes cell injury is not understood. Using a transgenic mouse model, we evaluated the effect of alpha-synuclein overexpression on gene expression in the substantia nigra. Nigral mRNA from wildtype and alpha-synuclein transgenic mice was analyzed using Affymetrix gene arrays. At three months, before pathological changes are apparent, we observed modest alterations in gene expression. However, nearly 200 genes were altered in expression at nine months, when degenerative changes are more apparent. Functional genomic analysis revealed that the genes altered at nine months were predominantly involved in gene transcription. As in human Parkinson's disease, gene expression changes in the transgenic model were also modulated by gender. These data demonstrate that alterations of gene expression are widespread in this animal model, and suggest that transcriptional dysregulation may be a disease mechanism that can be targeted therapeutically.

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Overexpression of human α-synuclein causes dopamine neuron death in primary human mesencephalic culture

Cited by 133 publications

References 41 publications

Effect of Mutant α-Synuclein on Dopamine Homeostasis in a New Human Mesencephalic Cell Line

Effect of Mutant α-Synuclein on Dopamine Homeostasis in a New Human Mesencephalic Cell Line

Inhibitory effect of c-Myc on p53-induced apoptosis in leukemia cells. Microarray analysis reveals defective induction of p53 target genes and upregulation of chaperone genes

Transcriptional dysregulation in a transgenic model of Parkinson disease

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