Speaker
Description
PD is characterized by the degeneration of dopaminergic neurons of the Substantia Nigra pars compacta (SNpc), disruption of dopamine (DA) homeostasis, and the formation of Lewy bodies (LB), intracellular protein inclusions primarily composed of α-synuclein (α-syn). Alpha-syn plays a key role in synaptic function regulating vesicle trafficking, membrane dynamics, and neurotransmitter release. The mechanisms underlying α-syn aggregation are not fully understood, but it is believed that α-syn undergoes a multistep aggregation process, forming toxic oligomers that disrupt DA homeostasis by damaging DA-containing vesicles. The SH-SY5Y cell line, derived from human neuroblastoma cells, is a commonly used model for studying neurodegenerative diseases like Parkinson’s disease (PD). However, differentiating SH-SY5Y cells into mature dopaminergic neurons remains a challenge due to the lack of a standardized protocol. Although several differentiation methods exist, their efficacy and reproducibility are often limited. To address this, we modified a previously proposed protocol to develop a more reliable and effective method for differentiating SH-SY5Y cells into dopaminergic neurons. This new protocol provides a robust model to investigate the molecular mechanisms underlying PD pathology. More specifically, we aimed at developing a cellular model to investigate how the interplay between DA and α-syn may guide the aggregation and spreading pathways. First, in-vitro α-syn aggregation studies were undertaken to explore the effects of DA-induced modifications on oligomers. Then, using our differentiated dopaminergic SH-SY5Y neurons, we investigated α-syn uptake, release, and cell-to-cell spreading in trans-well co-cultures exposed to fluorescently labelled recombinant α-syn protein monomers and oligomers. Eventually, we started the setup of methods to explore the effects of DA-induced modifications on α-syn seeding and spreading in three-dimensional cellular models (midbrain organoids). Collectively, our findings offer new insights into the formation and stabilization of α-syn oligomers in the presence of DA, contributing to a deeper understanding of the molecular mechanisms driving PD.
| Author(s) | Francesca Martorella*, Giorgia Pupo, Marta Lualdi, Tiziana Alberio, Mauro Fasano |
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| Affiliation(s) | Università degli Studi dell’Insubria, Dipartimento di Scienza e Alta Tecnologia, Busto Arsizio (VA) |
