Fig. 6

A53T SynGFP inclusions form in non-neuronal cells at longer intervals post-injection. a–c A53T SynGFP inclusions appear in at least three cell types: NeuN-positive cells (a), GFAP-positive cells (b), and Iba1-positive cells (c). The morphology and staining characteristics of these inclusions differed depending on cell-type identified via immunohistochemistry. For example, Iba1-positive SynGFP inclusions rarely co-stain with psyn (c). Scale bar 10 µm. d The percentage of A53T SynGFP inclusions in different cell types varies over time. At earlier intervals post-injection (less than 50 days post-injection (dpi)) A53T inclusions are more likely found in NeuN-positive cells than GFAP-positive cells or Iba1-positive cells, as identified via immunohistochemical analysis. At later intervals post-injection (greater than 50 dpi) A53T SynGFP inclusions are more likely found in GFAP-positive cells than Iba1-positive or NeuN-positive cells. Cells identified as Iba1-positive are unlikely to contain A53T SynGFP inclusions regardless of the post-injection interval, although a preference for the later interval is observed. NeuN = gray line, GFAP = green line, and Iba1 = blue line. e Group data shows that the cell-type dependence of A53T SynGFP inclusions varies in accordance with days post injection (dpi) (two-way ANOVA (interaction F(2,30) = 19.15), (Early vs. Late F(1, 30) = 0.9875), (cell type F(2,30) = 22.71), p < 0.0001, p = 0.3283, and p < 0.0001 respectively; Sidak’s multiple comparisons test, at early intervals there are more NeuN-positive inclusions bearing cells than at late intervals: p = 0.0029, at early intervals there are less GFAP-positive inclusion bearing cells than at late intervals: p < 0.0001, and there is no difference in the percentage Iba1-positive inclusions cells at the early and late timepoint: p = 0.9867; N = 6 animals per timepoint). Early intervals post-injection equal less than 50 days post-injection (dpi). Late intervals post-injection equal greater than 50 dpi. Mean and SEM of percentage of inclusions from each slice analyzed from each animal are shown at each time point. NeuN = gray, GFAP = green, and Iba1 = blue. f A53T SynGFP inclusion survival time is dependent on cell-type. In vivo imaging of individual cells over time showed variable rates of degeneration in inclusion-bearing cells. Based on morphology, neuronal and non-neuronal cell types were identified from in vivo data. Group data continues to show rapid degeneration of inclusions-bearing cells that are neuronal cells (median survival 7 days), however extended survival time is found in non-neuronal cells (data never reaches 50% survival); Mantel-cox test p < 0.0001. Neuronal cells = black, Non-neuronal cells = cyan