Fig. 4

Formation of α-syn inclusions impair spontaneous Ca²⁺ transients. Primary hippocampal neurons from wild type mice were exposed to fibrils (or PBS) at DIV 7. On DIV 8, primary neurons were transduced with AAV9-Synapsin1-GCamp6 to visualize Ca²⁺ transients selectively in neurons. On DIV 14, (7 days post induction of inclusions with fibrils), neurons were imaged live. Images were captured every 300 msec for 5 min at 37 °C. a Images of neurons expressing AAV9-Synapsin1-GCamp6. Scale bar = 100 μm. b Representative images of Ca²⁺ transients (pseudocolored so red represents high levels of GCamp6 fluorescence and blue represents low levels of GCamp6 fluorescence.) c Representative traces of Ca²⁺ transients over time in control neurons and neurons with inclusions. d Custom Matlab scripts quantified the frequency of Ca²⁺ spikes and amplitude of the Ca²⁺ spikes. Cumulative frequency plots show frequency and amplitude of spikes in control neurons (blue) and neurons with inclusions (red). Data was collected from three independent experiments. **** p < 0.0001 by the Kolmogorov-Smirnov test