A Single Cell-Based System To investigate Mechanisms of Neurodegeneration
Center for Applied Medical Research (CIMA), University of Navarra
Our laboratory has developed a microscope based methodology to identify mechanisms of death by simultaneously measuring in single neurons multiple features of interest and determining predictors of neuronal death -longitudinal survival analysis. This approach was successfully applied to determine predictive factors of neuronal death on a Huntington's disease neuronal model. Additional neuronal models of Alzheimer's disease, Parkinson's disease and Amyotrophic Lateral Sclerosis have been developed to study molecular mechanisms of neurodegeneration. Our most recent study has been focused on neurotoxic alpha-synuclein mechanisms. We have developed a primary neuronal model overexpressing wild type and mutant aSyn versions in which we have applied longitudinal survival analysis. We have determined how pathological hallmarks linked to aSyn toxicity contribute to neuronal death, like alterations in aSyn expression and stability (measured by optical-pulse labelling), aggregation or serine 129 phosphorylation (PS129). Finally, we have set up a novel assay to score for cell autonomous and non-autonomous aSyn-dependent toxicity. Currently, we are evaluating the role of the unfolding protein response (UPR) activation in determining neuronal survival/death in these neuronal models by using specific fluorescent reporters and pharmacological modulators of this cellular pathway.
Host: Fabio Cavaliere