RA Differentiation Enhances Dopaminergic Features, Changes Redox Parameters, and Increases Dopamine Transporter Dependency in 6-Hydroxydopamine-Induced Neurotoxicity in SH-SY5Y Cells.
Research on Parkinson's disease (PD) and drug development is hampered by the lack of suitable human in vitro models that simply and accurately recreate the disease conditions. To counteract this, many attempts to differentiate cell lines, such as the human SH-SY5Y neuroblastoma, into dopaminergic neurons have been undertaken since they are easier to cultivate when compared with other cellular models. Here, we characterized neuronal features discriminating undifferentiated and retinoic acid (RA)-differentiated SH-SYSY cells and described significant differences between these cell models in 6-hydroxydopamine (6-OHDA) cytotoxicity. In contrast to undifferentiated cells, RA-differentiated SH-SY5Y cells demonstrated low proliferative rate and a pronounced neuronal morphology with high expression of genes related to synapse vesicle cycle, dopamine synthesis/degradation, and of dopamine transporter (DAT). Significant differences between undifferentiated and RA-differentiated SH-SY5Y cells in the overall capacity of antioxidant defenses were found; although RA-differentiated SH-SY5Y cells presented a higher basal antioxidant capacity with high resistance against H2O2insult, they were twofold more sensitive to 6-OHDA. DAT inhibition by 3α-bis-4-fluorophenyl-methoxytropane and dithiothreitol (a cell-permeable thiol-reducing agent) protected RA-differentiated, but not undifferentiated, SH-SY5Y cells from oxidative damage and cell death caused by 6-OHDA. Here, we demonstrate that undifferentiated and RA-differentiated SH-SY5Y cells are two unique phenotypes and also have dissimilar mechanisms in 6-OHDA cytotoxicity. Hence, our data support the use of RA-differentiated SH-SY5Y cells as an in vitro model of PD. This study may impact our understanding of the pathological mechanisms of PD and the development of new therapies and drugs for the management of the disease.
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil. fe.m.lopes@gmail.com.|Institute of Pharmaceutical Science, King's College London, 150 Stamford Street, London, SE1 9NH, UK. fe.m.lopes@gmail.com.
Fernanda M
FM
Lopes
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil.
Leonardo Lisbôa
LL
da Motta
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil.
Marco A
MA
De Bastiani
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil.
Bianca
B
Pfaffenseller
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil.
Bianca W
BW
Aguiar
Cellular Defenses Laboratory, Department of Biochemistry, Biological Sciences Centre, Federal University of Santa Catarina (UFSC), Florianopolis, SC, 88040-900, Brazil.
Luiz F
LF
de Souza
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil.|Department of Physics, Federal University of Ceará (UFC), Fortaleza, CE, 60455-760, Brazil.
Geancarlo
G
Zanatta
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil.
Daiani M
DM
Vargas
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil.
Patrícia
P
Schönhofen
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil.
Giovana F
GF
Londero
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil.
Liana M
LM
de Medeiros
Department of Physics, Federal University of Ceará (UFC), Fortaleza, CE, 60455-760, Brazil.
Valder N
VN
Freire
Cellular Defenses Laboratory, Department of Biochemistry, Biological Sciences Centre, Federal University of Santa Catarina (UFSC), Florianopolis, SC, 88040-900, Brazil.
Alcir L
AL
Dafre
Bioinformatics and Systems Biology Laboratory, Polytechnic Center, Federal University of Paraná (UFPR), Curitiba, PR, 81520-260, Brazil.
Mauro A A
MA
Castro
Institute of Pharmaceutical Science, King's College London, 150 Stamford Street, London, SE1 9NH, UK.
Richard B
RB
Parsons
Laboratory of Cellular Biochemistry, Department of Biochemistry, ICBS/UFRGS, 2600 Ramiro Barcelos St, Porto Alegre, RS, 90035-003, Brazil. fabio.klamt@ufrgs.br.
Fabio
F
Klamt
D000975:Antioxidants / Q000378:metabolism*; D016923:Cell Death / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D004229:Dithiothreitol / Q000494:pharmacology; D050483:Dopamine Plasma Membrane Transport Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D006861:Hydrogen Peroxide; D010084:Oxidation-Reduction / Q000187:drug effects; D016627:Oxidopamine / Q000037:antagonists & inhibitors; D010720:Phosphines / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology*
D000975:Antioxidants; D004229:Dithiothreitol; D050483:Dopamine Plasma Membrane Transport Proteins; D006861:Hydrogen Peroxide; D016627:Oxidopamine; D010720:Phosphines; D014212:Tretinoin; C080938:tris(2-carboxyethyl)phosphine
6-hydroxydopamine; Dopamine transporter; Experimental model; Parkinson’s disease; Retinoic acid; SH-SY5Y cells