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Parkinson's disease (PD), adult neurodegenerative disorder, is caused by the death of dopamine neurons in the substantia nigra. High content screening (HCS) should allow finding new pathways involved in the onset of PD by screening molecules based on death phenotype. Rotenone, a chemical compound commonly used as pesticide, is well-documented as death inducer of dopamine neurons in the substantia nigra and allow to mimic PD in vitro and in vivo. Rotenone-induced degeneration of dopaminergic neurons may not be solely attributed to an impairment of neuronal mitochondrial complex I activity in the dopaminergic neurons but may also be boosted by the participation of the resident immune cells in the brain: the microglia cells. Effectively, various environmental factors may also work in concert to induce dopaminergic neurodegeneration and numerous studies have confirmed that neuroinflammation plays a critical role in the pathogenesis of neurodegenerative deseases, including PD.
To develop this assay, human neuroblastoma and mouse macrophagic cell lines in coculture were used, and rotenone was chosen as death inducer and phenotypic markers as Hoechst for nucleus and MAP2 for neurites analysis.
Furthermore, first assays in 3D cell co-culture show promising results and give us nice perspectives for the future.