Journal
FOOD AND CHEMICAL TOXICOLOGY
Volume 149, Issue -, Pages -Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.fct.2021.111967
Keywords
SH-SY5Y; Differentiation; Retinoic acid; Mitochondria; Neurotoxicology; Mitochondrial dysfunction
Categories
Funding
- Montepio Foundation
- FEDER/COMPETE/national funds by FCT [PTDC/BTMSAL/29297/2017, POCI-01-0145-FEDER-029297, UIDB/04539/2020, PTDC/MED-FAR/29391/2017, POCI-01-0145-FEDER-029391]
- ERDF through COMPETE 2020/FCT [PD/BD/128254/2016]
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A fast and reproducible in vitro model using differentiated SH-SY5Y cells was optimized to study mitochondria-related neurodegeneration, showing increased susceptibility to mitochondrial toxicants and potential for studying neurotoxicity processes, mitochondrial dynamics, and bioenergetic impairment.
Since most models used to study neuronal dysfunction display disadvantages and ethical concerns, a fast and reproducible in vitro model to study mitochondria-related neurodegeneration is required. Here, we optimized and characterized a 3-day retinoic acid-based protocol to differentiate the SH-SY5Y cell line into a neuronal-like phenotype and investigated alterations in mitochondrial physiology and distribution. Differentiation was associated with p21-linked cell cycle arrest and an increase in cell mass and area, possibly associated with the development of neurite-like extensions. Notably, increased expression of mature neuronal markers (neuronal-specific nuclear protein, microtubule-associated protein 2, beta III tubulin and enolase 2) was observed in differentiated cells. Moreover, increased mitochondrial content and maximal area per cell suggests mitochondrial remodeling. To demonstrate that this model is appropriate to study mitochondrial dysfunction, cells were treated for 6 h with mitochondrial toxicants (rotenone, antimycin A, carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP) and 6-hydroxydopamine (6-OHDA)). Differentiated cells were more susceptible to increasing concentrations of FCCP, antimycin A, and rotenone, while 6-OHDA showed a distinct dose-dependent neurotoxicity pattern. Even though differentiated cells did not exhibit a fully mature/differentiated neuronal phenotype, the protocol developed can be used to study neurotoxicity processes, mitochondrial dynamics, and bioenergetic impairment, representing an alternative to study mitochondrial impairment-related pathologies in vitro.
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