Comparison between proliferative and neuron-like SH-SY5Y cells as an in vitro model for Parkinson disease studies

• Published in: Brain research Vol. 1337; pp. 85 - 94
• Main Authors: Lopes, Fernanda Martins, Schröder, Rafael, Júnior, Mário Luiz Conte da Frota, Zanotto-Filho, Alfeu, Müller, Carolina Beatriz, Pires, André Simões, Meurer, Rosalva Thereza, Colpo, Gabriela Delevati, Gelain, Daniel Pens, Kapczinski, Flávio, Moreira, José Cláudio Fonseca, Fernandes, Marilda da Cruz, Klamt, Fabio
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 14-06-2010; Elsevier
• Subjects: 6-hydroxydopamine; Adrenergic Agents - toxicity; Adult and adolescent clinical studies; Animals; Biological and medical sciences; Biomarkers - analysis; Cattle; Cell Differentiation - drug effects; Cell Line, Tumor; Cell Proliferation - drug effects; Cell Survival - drug effects; Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases; DJ-1; Humans; Intracellular Signaling Peptides and Proteins - analysis; Keratolytic Agents - pharmacology; Medical sciences; Membrane Potential, Mitochondrial - drug effects; Mitochondria - drug effects; Neuroblastoma - metabolism; Neuroblastoma - pathology; Neurology; Oncogene Proteins - analysis; Oncogene Proteins - biosynthesis; Organic mental disorders. Neuropsychology; Oxidopamine - toxicity; Parkinson Disease; Parkinson Disease - metabolism; Parkinson Disease - pathology; Protein Deglycase DJ-1; Psychology. Psychoanalysis. Psychiatry; Psychopathology. Psychiatry; Tretinoin - pharmacology; Parkinson Disease; 6-hydroxydopamine; DJ-1; Nervous system diseases; Parkinson disease; Oxidopamine; In vitro; Cerebral disorder; Neuron; Central nervous system disease; Degenerative disease; Models; Comparative study; Extrapyramidal syndrome
• ISSN: 0006-8993; 1872-6240
• DOI: 10.1016/j.brainres.2010.03.102

Abstract The molecular mechanisms underlying the cellular lost found in the nigrostriatal pathway during the progression of Parkinson's disease (PD) are not completely understood. Human neuroblastoma cell line SH-SY5Y challenged with 6-hydroxydopamine (6-OHDA) has been widely used as an in vitro model for PD. Although this cell line differentiates to dopaminergic neuron-like cells in response to low serum and retinoic acid (RA) treatment, there are few studies investigating the differences between proliferative and RA-differentiated SH-SY5Y cells. Here we evaluate morphological and biochemical changes which occurs during the differentiation of SH-SY5Y cells, and their responsiveness to 6-OHDA toxicity. Exponentially growing SH-SY5Y cells were maintained with DMEM/F12 medium plus 10% of fetal bovine serum (FBS). Differentiation was triggered by the combination of 10 µM RA plus 1% of FBS during 4, 7 and 10 days in culture. We found that SH-SY5Y cells differentiated for 7 days show an increase immunocontent of several relevant neuronal markers with the concomitant decrease in non-differentiated cell marker. Moreover, cells became two-fold more sensitive to 6-OHDA toxicity during the differentiation process. Time course experiments showed loss of mitochondrial membrane potential triggered by 6-OHDA (mitochondrial dysfunction parameter), which firstly occurs in proliferative than neuron-like differentiated cells. This finding could be related to the increase in the immunocontent of the neuroprotective protein DJ-1 during differentiation. Our data suggest that SH-SY5Y cells differentiated by 7 days with the protocol described here represent a more suitable experimental model for studying the molecular and cellular mechanisms underlying the pathophysiology of PD.