Sebastian Illes, SwedenCellectricon AB CNS Research
Author Of 1 Presentation
IN VITRO MODELING OF SYNUCLEINOPATHY ON A HIGH CAPACITY MICROFLUIDICS PLATFORM
The appearance and spreading of misfolded alpha-synuclein are common neuropathological hallmarks in brain cells of patients suffering from Parkinson’s disease. Deciphering the mechanisms behind uptake, transport, spreading, aggregation, and propagation of alpha-synuclein is of high relevance for preclinical medical research programs. Using a high capacity assay platform for in vitro modelling of disease mechanisms, we aim to evaluate current hypotheses and test novel strategies for prevention of progressive synucleinopathy.
Mouse E18 cortical neurons were cultured in the wells of a microfluidic co-culture plate containing 96 experimental units. Different assay formats were developed for the monitoring of uptake, axonal transport and somatic accumulation of fluorescently labelled alpha-synuclein as well as aggregation of endogenous alpha-synuclein in cortical neurons. These individual neuropathological processes were assessed by live-cell imaging, immunocytochemistry and HCA on different timepoints up to 20 days in vitro.
Our data show that fluorescently labelled alpha-synuclein seeds are taken up by axons, are retrogradely transported and accumulate within the soma of cortical neurons. We demonstrate that cortical neurons filled with alpha-synuclein seeds cause aggregation of endogenous alpha-synuclein.
The assay formats presented are suitable for in vitro modelling of neuropathologies. In addition, the assay platform allows for intervention approaches where not only the prevention of pathology can be detected, but also the study of effects on specific disease mechanisms.
The presented platform shows sufficient capacity and robustness to allow for screening and profiling of larger compound sets, in the search for molecules preventing specific processes occurring in progressive synucleinopathy.