Neurotoxicity of “ecstasy” and its metabolites in human dopaminergic differentiated SH-SY5Y cells

Ferreira, Patrícia Silva; Nogueira, T.; Costa, Vera Marisa; Branco, Paula S.; Ferreira, Laura Fernandes; Fernandes, Eduarda; Bastos, Maria de Lourdes; Meisel, Andreas; Carvalho, Félix; Capela, João Paulo

doi:10.1016/j.toxlet.2012.11.015

Cited by 40 publications

(33 citation statements)

References 49 publications

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“…However, MDMA did not sensitize the cells to deleterious effects of high temperature. Although the concentrations of MDMA were high (up to 1 mM) they corresponded to the concentrations required to obtain cytotoxicity in other studies using other cell culture systems [3,55,56]. In our mouse cell model, exposure to 1 mM MDMA at 37°C for 72 hours produced a maximal cell death of 60% (as shown by the MTT data).…”

Section: Discussionmentioning

confidence: 47%

“…Glutamate excitotoxicity has also been implicated in MDMA toxicity in vivo [6,7], but this is unlikely to be the case in the P19 neurons, since glutamate produces only mild effects even at high concentrations [32]. Metabolites of MDMA have been suggested to be the major contributors for MDMA neurotoxicity in human cell lines [55], but this has not been addressed in the present study.…”

Section: Discussionmentioning

confidence: 94%

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Non-Serotonergic Neurotoxicity by MDMA (Ecstasy) in Neurons Derived from Mouse P19 Embryonal Carcinoma Cells

et al. 2016

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3,4-methylenedioxymethamphetamine (MDMA; ecstasy) is a commonly abused recreational drug that causes neurotoxic effects in both humans and animals. The mechanism behind MDMA-induced neurotoxicity is suggested to be species-dependent and needs to be further investigated on the cellular level. In this study, the effects of MDMA in neuronally differentiated P19 mouse embryonal carcinoma cells have been examined. MDMA produces a concentration-, time- and temperature-dependent toxicity in differentiated P19 neurons, as measured by intracellular MTT reduction and extracellular LDH activity assays. The P19-derived neurons express both the serotonin reuptake transporter (SERT), that is functionally active, and the serotonin metabolizing enzyme monoamine oxidase A (MAO-A). The involvement of these proteins in the MDMA-induced toxicity was investigated by a pharmacological approach. The MAO inhibitors clorgyline and deprenyl, and the SERT inhibitor fluoxetine, per se or in combination, were not able to mimic the toxic effects of MDMA in the P19-derived neurons or block the MDMA-induced cell toxicity. Oxidative stress has been implicated in MDMA-induced neurotoxicity, but pre-treatment with the antioxidants α-tocopherol or N-acetylcysteine did not reveal any protective effects in the P19 neurons. Involvement of mitochondria in the MDMA-induced cytotoxicity was also examined, but MDMA did not alter the mitochondrial membrane potential (ΔΨm) in the P19 neurons. We conclude that MDMA produce a concentration-, time- and temperature-dependent neurotoxicity and our results suggest that the mechanism behind MDMA-induced toxicity in mouse-derived neurons do not involve the serotonergic system, oxidative stress or mitochondrial dysfunction.

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Section: Discussionmentioning

confidence: 47%

Section: Discussionmentioning

confidence: 94%

Non-Serotonergic Neurotoxicity by MDMA (Ecstasy) in Neurons Derived from Mouse P19 Embryonal Carcinoma Cells

et al. 2016

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“…Through differentiation (see protocol below), SH-SY5Y cells are able to express dopaminergic markers, as well as tyrosine hydroxylase, DAT, and higher ability to accumulate dopamine and exhibit extended neurites [17, 18]. In this way, the cells are no longer immature, mitosis rate is reduced, and also differentiation agents can mimic factors secreted by astrocytes in a the natural brain environment.…”

Section: Methodsmentioning

confidence: 99%

Differential Effects of Methyl-4-Phenylpyridinium Ion, Rotenone, and Paraquat on Differentiated SH-SY5Y Cells

Martins

Bastos

Carvalho

et al. 2013

Journal of Toxicology

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Paraquat (PQ), a cationic nonselective bipyridyl herbicide, has been used as neurotoxicant to modulate Parkinson's disease in laboratory settings. Other compounds like rotenone (ROT), a pesticide, and 1-methyl-4-phenylpyridinium ion (MPP+) have been widely used as neurotoxicants. We compared the toxicity of these three neurotoxicants using differentiated dopaminergic SH-SY5Y human cells, aiming to elucidate their differential effects. PQ-induced neurotoxicity was shown to be concentration and time dependent, being mitochondrial dysfunction followed by neuronal death. On the other hand, cells exposure to MPP+ induced mitochondrial dysfunction, but not cellular lyses. Meanwhile, ROT promoted both mitochondrial dysfunction and neuronal death, revealing a biphasic pattern. To further elucidate PQ neurotoxic mechanism, several protective agents were used. SH-SY5Y cells pretreatment with tiron (TIR) and 2-hydroxybenzoic acid sodium salt (NaSAL), both antioxidants, and N ω-nitro-L-arginine methyl ester hydrochloride (L-NAME), a nitric oxide synthase inhibitor, partially protected against PQ-induced cell injury. Additionally, 1-(2-[bis(4-fluorophenyl)methoxy]ethyl)-4-(3-phenyl-propyl)piperazine (GBR 12909), a dopamine transporter inhibitor, and cycloheximide (CHX), a protein synthesis inhibitor, also partially protected against PQ-induced cell injury. In conclusion, we demonstrated that PQ, MPP+, and ROT exerted differential toxic effects on dopaminergic cells. PQ neurotoxicity occurred through exacerbated oxidative stress, with involvement of uptake through the dopamine transporter and protein synthesis.

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“…A more mature neuronal phenotype can be reached using growth medium supplemented with specific chemical factors, such as recombinant human brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), dibutyryl-cyclic AMP (db-cAMP), phorbol esters and others (Agholme et al 2010;Dwane et al 2013;Encinas et al 2000;Gimenez-Cassina et al 2006;Jamsa et al 2004;Kou et al 2008;Kume et al 2008;Sarkanen et al 2007). Thereafter, SH-SY5Y cells have been largely utilized to investigate the disease mechanisms associated with various neurodegenerative disorders (Agholme et al 2014;Ferreira et al 2013;Hadzhieva et al 2013;Jamsa et al 2004;Ke et al 2012;Krishna et al 2014;Lim et al 2015;Lopes et al 2010Lopes et al , 2012Palomo et al 2011;Scifo et al 2013Scifo et al , 2015.…”

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confidence: 99%

Transcriptomic Profiling Discloses Molecular and Cellular Events Related to Neuronal Differentiation in SH-SY5Y Neuroblastoma Cells

et al. 2016

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Human SH-SY5Y neuroblastoma cells are widely utilized in in vitro studies to dissect out pathogenetic mechanisms of neurodegenerative disorders. These cells are considered as neuronal precursors and differentiate into more mature neuronal phenotypes under selected growth conditions. In this study, in order to decipher the pathways and cellular processes underlying neuroblastoma cell differentiation in vitro, we performed systematic transcriptomic (RNA-seq) and bioinformatic analysis of SH-SY5Y cells differentiated according to a two-step paradigm: retinoic acid treatment followed by enriched neurobasal medium. Categorization of 1989 differentially expressed genes (DEGs) identified in differentiated cells functionally linked them to changes in cell morphology including remodelling of plasma membrane and cytoskeleton, and neuritogenesis. Seventy-three DEGs were assigned to axonal guidance signalling pathway, and the expression of selected gene products such as neurotrophin receptors, the functionally related SLITRK6, and semaphorins, was validated by immunoblotting. Along with these findings, the differentiated cells exhibited an ability to elongate longer axonal process as assessed by the neuronal cytoskeletal markers biochemical characterization and morphometric evaluation. Recognition of molecular events occurring in differentiated SH-SY5Y cells is critical to accurately interpret the cellular responses to specific stimuli in studies on disease pathogenesis.

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Neurotoxicity of “ecstasy” and its metabolites in human dopaminergic differentiated SH-SY5Y cells

Cited by 40 publications

References 49 publications

Non-Serotonergic Neurotoxicity by MDMA (Ecstasy) in Neurons Derived from Mouse P19 Embryonal Carcinoma Cells

Non-Serotonergic Neurotoxicity by MDMA (Ecstasy) in Neurons Derived from Mouse P19 Embryonal Carcinoma Cells

Differential Effects of Methyl-4-Phenylpyridinium Ion, Rotenone, and Paraquat on Differentiated SH-SY5Y Cells

Transcriptomic Profiling Discloses Molecular and Cellular Events Related to Neuronal Differentiation in SH-SY5Y Neuroblastoma Cells

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