Nathalie Maubon, Marie-Christine Bret, Christelle Odrobina, Philippe Masson, and Mireille Tallandier. (Abbott Group, Laboratoires Fournier, Research Center, Global System Biology Department, HCS unit, 50 rue de Dijon, 21121 DAIX, France)
Development of a High Throughput Neuroprotection Assay using High Content Screening Technology for Treatment Research in Parkinson's Disease.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. To develop this assay, human neuroblastoma SH-SY5Y cell line, rotenone as death inducer and death markers were used. Hoechst was used as nucleus marker. Annexin V served as apoptotic marker with recognition of phosphatidyl-serine flip-flop during apoptosis. Propidium iodide, which enters the cells only when the membrane is disrupted, was used as necrotic marker. Dose effect of rotenone and Z score analysis allowed determining the rotenone concentration used in this model at 3 µM. SH-SY5Y cell line with rotenone at 3 µM showed around 70% of cell death, Z-scores on Annexin V analysis reached 0.35 and on DNA condensation around 0.5. Propidium iodide allowed differentiating between early and late apoptosis. Some pathway inhibitors were assayed in this set-up and showed slight neuroprotection effect. In conclusion, this HCS could allow us to identify new compounds with neuroprotective effect using new pathways for PD treatment.
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