Fig. 6
PD-type inclusion formation in AAV-hTyr-injected rats. a Quantification of NM-laden neurons with p62-positive intranuclear Marinesco bodies (MB) in ipsilateral SNpc from AAV-hTyr-injected rats. *p < 0.05, compared to 0.5 m (ANOVA on ranks; Dunn’s post-hoc test). b Quantification of NM-laden neurons with p62-positive total intracytoplasmic inclusion bodies (left) and pale bodies (PB) and/or Lewy body (LB)-like p62-positive inclusions (right) in ipsilateral SNpc from AAV-hTyr-injected rats. Left, *p < 0.05, compared to 0.5, 1, 4, 12, and 24 m (one-way ANOVA; Student–Newman–Keuls post-hoc test). Right, *p < 0.05, compared to PB at 0.5, 4, 12, and 24 m; #p < 0.05, compared to LB-like inclusions at 0.5, 1, 4, and 24 m (two-way ANOVA; Holm-Sidak post-hoc test). c–e Midbrain sections from PD post-mortem brains (top) and AAV-hTyr-injected rats (bottom) exhibiting MB (c, white arrowheads), PB (d, blue arrowheads) and LB-type aSyn-positive inclusions (e, arrows). In c, d, Hematoxylin-eosin (H&E) staining. In e, aSyn immunostaining (in blue). Scale bars, 12.5 μm. f Ipsilateral SNpc sections from AAV-hTyr-injected rats exhibiting NM-laden neurons with MB (white arrowhead) detected by immunofluorescence with p62 (red) and ubiquitin (Ub, green). Nuclei are stained with Hoechst (blue). Scale bar, 12.5 μm. g Ipsilateral SNpc sections from AAV-hTyr-injected rats exhibiting NM-laden neurons with p62-positive multipunctate cytoplasmic inclusions (yellow arrowhead), PB (blue arrowhead) or LB-like inclusions (arrows). Scale bar, 12.5 μm. h Ipsilateral SNpc section from an AAV-hTyr-injected rat exhibiting a NM-laden neuron with an intracytoplasmic LB-like inclusion immunopositive for p62 (red), aSyn (purple) and ubiquitin (Ub, green). Scale bar, 12.5 μm. i Ipsilateral SNpc section from an AAV-hTyr-injected rat immunostained for p62 (red) and TH (purple) in which a NM-laden neuron containing both a PB (blue arrowhead) and a MB (white arrowhead) exhibits decreased TH immunostaining (asterisk). Nuclei are stained with Hoechst (blue). Scale bar, 25 μm. In a, b, values are mean ± SEM. n = 8 (0.5 m), n = 5 (1 m), n = 6 (2 m), n = 5 (4 m), n = 6 (12 m), n = 5 (24 m). Ub ubiquitin, BF bright-field. In all panels, unstained NM appears as brown. Photomicrographs correspond to 5-μm-thick sections
