Proteomics analysis of MPP+-induced apoptosis in SH-SY5Y cells

Xie, Hongrong; Chen, Ming; Hu, Xinyu; Wang, Danping; Tian, Mingxiu; Li, Guoyi; Jiang, Hao-Bo; Wang, Ying; Dong, Zhong; Zhang, Yuhua; Hu, Linsen

doi:10.1007/s10072-010-0340-3

Cited by 18 publications

(21 citation statements)

References 24 publications

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“…Thus, exposure of cells to toxins that mimic cell death mechanisms associated with PD for short periods (3 and 24 h) is a useful model for studying the early events which take place when cells are under cell stress. Given that catecholaminergic neurons are the most susceptible to neurodegeneration in PD (Hornykiewicz 1972;Braak et al 1996), the catecholaminergic neuroblastoma cell line, SH-SY5Y, which has previously been used as a model for studying cell death mechanisms in PD was selected for our studies (Nonaka and Hasegawa 2009;Wu et al 2009;Xie et al 2010Xie et al , 2011Dadakhujaev et al 2010). Whilst primary mesencephalic cultures are a more physiological representation of neurons of the substantia nigra pars compacta, it is difficult to produce them in large quantities, and so it Western blots to show changes in ubiquitin levels following toxin exposure.…”

Section: Discussionmentioning

confidence: 99%

“…Here we sought to identify such a point of convergence by using a catecholaminergic neuroblastoma cell line, which has previously been used to study the mechanisms underlying cell death in PD (Nonaka and Hasegawa 2009;Wu et al 2009;Xie et al 2010Xie et al , 2011Dadakhujaev et al 2010), in combination with toxins that induce cell death via the three mechanisms commonly associated with neurodegeneration in PD: inhibition of mitochondrial complex I (rotenone), inhibition of the UPS using Z-Ile-Glu(OBu t )-Ala-Leu-H (PSI), and disruption of the lysosomal membrane via 5, 8-dihydroxy-1,4-naphthoquinone (naphthazarin). In order to determine the effect of these toxins in early, as compared to later stages of cell stress, when cell damage is unlikely to be reversible, sub-cellular function was measured after 3 and 24 h of exposure to toxin.…”

Section: Introductionmentioning

confidence: 99%

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Mitochondrial Dysfunction Precedes Other Sub-Cellular Abnormalities in an In Vitro Model Linked with Cell Death in Parkinson’s Disease

et al. 2011

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Dysfunction of mitochondria, the ubiquitin proteasome system (UPS), and lysosomes are believed to contribute to the pathogenesis of Parkinson's disease (PD). If it were possible to rescue functionally compromised, but still viable neurons early in the disease process, this would slow the rate of neurodegeneration. Here, we used a catecholaminergic neuroblastoma cell line (SH-SY5Y) as a model of susceptible neurons in PD. To identify a target early in the cell death process that was common to all neurodegenerative processes linked with PD, cells were exposed to toxins that mimic cell death mechanisms associated with PD. The sub-cellular abnormalities that occur shortly after toxin exposure were determined. 3 h of exposure to either naphthazarin, to inhibit lysosomal function, Z-Ile-Glu(OBu(t))-Ala-Leu-H (PSI), to inhibit the UPS, or rotenone, to inhibit mitochondrial complex I, caused depolarisation of the mitochondrial membrane potential (2.5-fold, twofold, and 4.6-fold change, respectively compared to vehicle), suggesting impaired mitochondrial function. Following 24 h exposure to the same toxins, UPS and lysosomal function were also impaired, and ubiquitin levels were increased. Thus, following exposure to toxins that mimic three important, but disparate cell death mechanisms associated with PD, catecholaminergic cells initially experience mitochondrial dysfunction, which is then followed by abnormalities in UPS and lysosomal function. Thus, mitochondrial dysfunction is an early event in cell stress. We suggest that, in patients with PD, the surviving cells of the substantia nigra pars compacta are most susceptible to mitochondrial impairment. Thus, targeting the mitochondria may be useful for slowing the progression of neurodegeneration in PD.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Mitochondrial Dysfunction Precedes Other Sub-Cellular Abnormalities in an In Vitro Model Linked with Cell Death in Parkinson’s Disease

et al. 2011

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“…Overexpression or application of S3 protein leads to improved neuroprotection against experimental cerebral ischemic damage and against cell death in a mouse model of Parkinson's disease (Ahn et al, ; Hwang et al, ). Similarly, levels of P0 protein were increased in an in vitro model of Parkinson's disease and have thus been hypothesized to be neuroprotective (Xie et al, ). Because there is no evidence that PrP directly interacts with S3 and P0, enhanced exosomal levels of these proteins are unlikely due to their increased exosomal packaging via association to PrP.…”

Section: Discussionmentioning

confidence: 99%

Improvement of neuronal cell survival by astrocyte‐derived exosomes under hypoxic and ischemic conditions depends on prion protein

Guitart

Loers

Buck

et al. 2016

Glia

125

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Prion protein (PrP) protects neural cells against oxidative stress, hypoxia, ischemia, and hypoglycemia. In the present study we confirm that cultured PrP-deficient neurons are more sensitive to oxidative stress than wild-type neurons and present the novel findings that wild-type, but not PrP-deficient astrocytes protect wild-type cerebellar neurons against oxidative stress and that exosomes released from stressed wild-type, but not from stressed PrP-deficient astrocytes reduce neuronal cell death induced by oxidative stress. We show that neuroprotection by exosomes of stressed astrocytes depends on exosomal PrP but not on neuronal PrP and that astrocyte-derived exosomal PrP enters into neurons, suggesting neuronal uptake of astrocyte-derived exosomes. Upon exposure of wild-type astrocytes to hypoxic or ischemic conditions PrP levels in exosomes were increased. By mass spectrometry and Western blot analysis, we detected increased levels of 37/67 kDa laminin receptor, apolipoprotein E and the ribosomal proteins S3 and P0, and decreased levels of clusterin/apolipoprotein J in exosomes from wild-type astrocytes exposed to oxygen/glucose deprivation relative to exosomes from astrocytes maintained under normoxic conditions. The levels of these proteins were not altered in exosomes from stressed PrP-deficient astrocytes relative to unstressed PrP-deficient astrocytes. These results indicate that PrP in astrocytes is a sensor for oxidative stress and mediates beneficial cellular responses, e.g. release of exosomes carrying PrP and other molecules, resulting in improved survival of neurons under hypoxic and ischemic conditions.

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“…Even though the effects of MeHg or MPP + on protein or gene expression patterns in the context of neurotoxicity have been reported in previous studies (Keyvanshokooh et al, 2009;Vendrell et al, 2010;Xie et al, 2011), this is the first report to use both genomic and proteomic approach to study the effects of MeHg and its relationship with PD using a dopaminergic cell line. Using a known shown to be involved in PD pathogenesis.…”

Section: Discussionmentioning

confidence: 99%

Methylmercury can induce Parkinson’s-like neurotoxicity similar to 1-methyl-4- phenylpyridinium: a genomic and proteomic analysis on MN9D dopaminergic neuron cells

et al. 2015

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-Exposure to environmental chemicals has been implicated as a possible risk factor for the development of neurodegenerative diseases. Our previous study showed that methylmercury (MeHg) exposure can disrupt synthesis, uptake and metabolism of dopamine similar to 1-methyl-4-phenylpyridinium (MPP + ). The objective of this study was to investigate the effects of MeHg exposure on gene and protein profiles in a dopaminergic MN9D cell line. MN9D cells were treated with MeHg (1-5 μM) and MPP + (10-40 μM) for 48 hr. Real-time PCR Parkinson's disease (PD) arrays and highperformance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/ MS) were performed for the analysis. PD PCR array results showed that 19% genes were significantly changed in the 2.5 μM MeHg treated cells, and 39% genes were changed in the 5 μM MeHg treated cells. In comparison, MPP + treatment (40 μM) resulted in significant changes in 25% genes. A total of 15 common genes were altered by both MeHg and MPP + , and dopaminergic signaling transduction was the most affected pathway. Proteomic analysis identified a total of 2496 proteins, of which 188, 233 and 395 proteins were differentially changed by 1 μM and 2.5 μM MeHg, and MPP + respectively. A total of 61 common proteins were changed by both MeHg and MPP + treatment. The changed proteins were mainly involved in energetic generation-related metabolism pathway (propanoate metabolism, pyruvate metabolism and fatty acid metabolism), oxidative phosphorylation, proteasome, PD and other neurodegenerative disorders. A total of 7 genes/proteins including Ube2l3 (Ubiquitin-conjugating enzyme E2 L3) and Th (Tyrosine 3-monooxygenase) were changed in both genomic and proteomic analysis. These results suggest that MeHg and MPP + share many similar signaling pathways leading to the pathogenesis of PD and other neurodegenerative diseases.

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Proteomics analysis of MPP+-induced apoptosis in SH-SY5Y cells

Cited by 18 publications

References 24 publications

Mitochondrial Dysfunction Precedes Other Sub-Cellular Abnormalities in an In Vitro Model Linked with Cell Death in Parkinson’s Disease

Mitochondrial Dysfunction Precedes Other Sub-Cellular Abnormalities in an In Vitro Model Linked with Cell Death in Parkinson’s Disease

Improvement of neuronal cell survival by astrocyte‐derived exosomes under hypoxic and ischemic conditions depends on prion protein

Methylmercury can induce Parkinson’s-like neurotoxicity similar to 1-methyl-4- phenylpyridinium: a genomic and proteomic analysis on MN9D dopaminergic neuron cells

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