title: Comparison between proliferative and neuron-like SH-SY5Y cells as an in vitro model for Parkinson disease studies.
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 microM 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.
authors.affiliation: Department of Biochemistry, ICBS/UFRGS, Porto Alegre/RS, Brazil.
authors.forename: Fernanda Martins
authors.initials: FM
authors.lastname: Lopes
authors.forename: Rafael
authors.initials: R
authors.lastname: Schröder
authors.forename: Mário Luiz Conte
authors.initials: ML
authors.lastname: da Frota
authors.forename: Alfeu
authors.initials: A
authors.lastname: Zanotto-Filho
authors.forename: Carolina Beatriz
authors.initials: CB
authors.lastname: Müller
authors.forename: André Simões
authors.initials: AS
authors.lastname: Pires
authors.forename: Rosalva Thereza
authors.initials: RT
authors.lastname: Meurer
authors.forename: Gabriela Delevati
authors.initials: GD
authors.lastname: Colpo
authors.forename: Daniel Pens
authors.initials: DP
authors.lastname: Gelain
authors.forename: Flávio
authors.initials: F
authors.lastname: Kapczinski
authors.forename: José Cláudio Fonseca
authors.initials: JC
authors.lastname: Moreira
authors.forename: Marilda da Cruz
authors.initials: Mda C
authors.lastname: Fernandes
authors.forename: Fabio
authors.initials: F
authors.lastname: Klamt
mesh_terms: D018663:Adrenergic Agents / Q000633:toxicity*; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis; D007641:Keratolytic Agents / Q000494:pharmacology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology*; D015513:Oncogene Proteins / Q000032:analysis / Q000096:biosynthesis; D016627:Oxidopamine / Q000633:toxicity*; D010300:Parkinson Disease / Q000378:metabolism / Q000473:pathology*; D000071617:Protein Deglycase DJ-1; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D018663:Adrenergic Agents; D015415:Biomarkers; D047908:Intracellular Signaling Peptides and Proteins; D007641:Keratolytic Agents; D015513:Oncogene Proteins; D014212:Tretinoin; D016627:Oxidopamine; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1