Proteomics as a tool to investigate cell models for dopamine toxicity

Fasano, Mauro; Alberio, Tiziana; Colapinto, Monica; Mila, Silvia; Lopiano, Leonardo

doi:10.1016/j.parkreldis.2008.04.016

Cited by 13 publications

(12 citation statements)

References 31 publications

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“…For example, SH-SY5Y neuroblastoma cells, used as a model for Parkinson's disease [47] and dopamine toxicity [48], can have several distinct phenotypes. This cell line has been used extensively in adherent nondifferentiated or differentiated states in many studies.…”

Section: Pepp0 As a Probe For Assessing Intracellular O 2 Levelsmentioning

confidence: 99%

Bactenecin 7 peptide fragment as a tool for intracellular delivery of a phosphorescent oxygen sensor

et al. 2010

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Research on cell‐penetrating peptides for the intracellular delivery of porphyrin compounds has mainly focused on the use of trans‐activator of transcription (TAT)‐derived peptides and, to a lesser extent, on proline‐rich peptides and phosphorescent metalloporphyrins. In this article, we describe a novel phosphorescent oxygen‐sensitive probe for intracellular use which comprises a bactenecin 7 peptide fragment (15–24) conjugated with the uncharged monofunctional derivative of Pt(II) coproporphyrin I (PEPP0). This probe provides efficient loading of various mammalian cells, including PC12, HCT116, SH‐SY5Y and HeLa, via cell‐type‐dependent uptake mechanisms. The conjugate displays a similar distribution in cytoplasm and mitochondria which allows local oxygen levels to be monitored. Respiratory responses of PC12 cells loaded with the conjugate, measured on a time‐resolved fluorescent reader, showed significant cell deoxygenation in response to uncoupling by carbonyl cyanide 4‐(trifluoromethoxy)phenylhydrazone and external hypoxia. Treatment with mitochondrial inhibitors led to a decrease in cell deoxygenation. Although the biophysical properties of this conjugate are similar to those of the phosphorescent intracellular oxygen‐sensitive probes described previously, it possesses a number of advantages, including ease of synthesis, high loading efficiency and reliability in physiological experiments with cells. This intracellular probe can be employed for the measurement of intracellular O2 levels in samples containing mammalian cells using the phosphorescence quenching technique. In addition, the responses to metabolic stimuli can be assessed in a wide range of cells, as can the levels of relative cell oxygenation under external hypoxia.

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Section: Pepp0 As a Probe For Assessing Intracellular O 2 Levelsmentioning

confidence: 99%

Bactenecin 7 peptide fragment as a tool for intracellular delivery of a phosphorescent oxygen sensor

et al. 2010

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“…The SH-SY5Y human neuroblastoma cell line is widely used as a cellular model to mimic impaired dopamine homeostasis (8,9,(18)(19)(20). In this cell line, we measured cell viability and apoptosis after 24 h exposure to different dopamine concentrations.…”

Section: Resultsmentioning

confidence: 99%

“…For two-dimensional electrophoresis, 3.5 3 10 6 cells were seeded in 3% FBS and treated with 0.1 mM dopamine in the presence of 700 U/ mL catalase in the medium to remove exogenous H 2 O 2 (8,9). After 24 h incubation, cells were scraped, collected by centrifugation, and lysed with 200 lL UTC (7 M urea, 2 M thiourea, 4% w/v 3-[(3-cholamido)dimethylammonio]-1-propanesulfonic acid (CHAPS)) containing protease inhibitors, and sonicated for 15 sec.…”

Section: Cell Lines and Culture Conditionsmentioning

confidence: 99%

“…Nigral dopaminergic neurons are particularly exposed to oxidative stress because the metabolism of dopamine gives rise to various molecules that can act as endogenous toxins if not handled properly (1)(2)(3)(4)(5). Homeostasis of ROS arising from altered dopamine compartmentalization is actively regulated by DJ-1, a peroxiredoxin-like antioxidant enzyme (6)(7)(8)(9)(10). Two homozygous mutations in the DJ-1 gene that prevent the expression of DJ-1 protein were associated to a familial form of early-onset autosomal recessive PD (PARK7), thus suggesting that loss of DJ-1 function leads to neurodegeneration (11).…”

Section: Introductionmentioning

confidence: 99%

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Changes in the two‐dimensional electrophoresis pattern of the Parkinson's disease related protein DJ‐1 in human SH‐SY5Y neuroblastoma cells after dopamine treatment

Alberio

Colapinto

Natale³

et al. 2010

IUBMB Life

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SummaryDJ-1 is a mitochondrial protein linked to Parkinson's disease. DJ-1 has been suggested to have several possible functions, although it has been mainly associated to oxidative stress defence. Changes in the two-dimensional electrophoresis pattern have been thoroughly described as a consequence of oxidative modification of the Cys106 residue. There is accumulating evidence supporting a specific role of DJ-1 in protecting dopaminergic neurons from dopamine itself. By exposing SH-SY5Y human neuroblastoma catecholaminergic cells to dopamine, we observed a specific increase in the most acidic forms in the DJ-1 two-dimensional electrophoresis pattern together with a significant decrease of the most basic spot. Unlike cells exposed to generic oxidative conditions, no additional shift was observed. The results are corroborated by a meta-analysis of the literature showing that in the absence of dopamine treatment the specific acidic form is underrepresented. IUBMBIUBMB Life, 62(9): 688-692, 2010

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“…More reliable are the studies performed on protein aggregates characterizing the neurodegenerative conditions, like Lewy bodies, amyloid plaques, and neurofibrillary tangles (Cumming & Schubert, 2005;Leverenz et al, 2007;Liao et al, 2004;Wakabayashi et al, 2007;Wang et al, 2005) because these are more stable structures. Proteomics analysis of brain has been more frequently used when animal models of neurodegenerative diseases are investigated (Gotz & Ittner, 2008), while some studies have been performed on cellular models, principally cell lines as the dopaminergic neuroblastoma SH-SY5Y (Alberio et al, 2010;Fasano, Alberio, Colapinto, Mila, & Lopiano, 2008).…”

Section: Sample Tissuesmentioning

confidence: 99%

Comparative Proteomics for the Evaluation of Protein Expression and Modifications in Neurodegenerative Diseases

Conti

Alessio

2015

International Review of Neurobiology

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Proteomics as a tool to investigate cell models for dopamine toxicity

Cited by 13 publications

References 31 publications

Bactenecin 7 peptide fragment as a tool for intracellular delivery of a phosphorescent oxygen sensor

Bactenecin 7 peptide fragment as a tool for intracellular delivery of a phosphorescent oxygen sensor

Changes in the two‐dimensional electrophoresis pattern of the Parkinson's disease related protein DJ‐1 in human SH‐SY5Y neuroblastoma cells after dopamine treatment

Comparative Proteomics for the Evaluation of Protein Expression and Modifications in Neurodegenerative Diseases

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