Fig. 4
A53T SynGFP mice form Lewy Body inclusions at an expedited rate following PFF injection and show a rapid cell death time course. a–f In vivo imaging of individual cells over time shows rapid inclusion formation and degeneration of inclusion-bearing cells. Example images show an individual cell tracked over time (arrows). The cell was followed beginning at 6 dpi and shows SynGFP fluorescence in puncta at terminals and homogeneous fluorescence in the cell body indicating soluble SynGFP (white arrows). At 10 dpi the inclusion begins to form and SynGFP fluorescence starts to clear from the nucleus and become more intense at the edges of the cytoplasm, indicating aggregated SynGFP (green arrows). At 15 dpi SynGFP aggregation is clearly visible in the processes of the cell (arrow heads). Within a 2-day window the inclusion disappears and at 17 dpi the inclusion-bearing cell has degenerated (red arrow). Survival time of this inclusion-bearing cell was 7 days. Scale bar 10 µm. g–l A53T SynGFP inclusions form at a much faster rate compared to previously published data tracking inclusion formation in WT SynGFP mice. A total of six A53T SynGFP mice were tracked in this study (1–6). Individual cells were tracked overtime in cortex layer II/III of both the injected (ipsi) and un-injected (contra) hemispheres. In all mice mature SynGFP inclusions started forming within 10 days post injection (dpi) compared to the 3–4 months post injection (mpi) time frame previously reported in WT SynGFP mice. Formation rate was tracked as both a measure of overall density (inclusions/mm3) (g, i, k) and as a percentage of total inclusions formed (h, j, l). Individual mice (g, h), group data as a function of location (ipsi, contra) (i, j) and overall group data (k, l) and are shown. A53T SynGFP mice show both intra and inter-animal variability in the density and percentage of inclusions formed at each time point (SEM), but the overall rate of formation followed the same rapid time course in all mice. m Group data shows rapid degeneration of inclusion-bearing cells, with a median survival time of 8 days after inclusion formation