Figure S1. TH depletion and partial recovery in both SNpc and striatum following acute MPTP treatment in mice. TH-DAB immunostaining in the SNpc (AC) and striatum (DF) of control (A,D) and MPTP-injected mice (B,C,E,F). (A,D) In control animals, TH immunolabelling is intense in both cell bodies and axons of the dopaminergic neurones. (B,E) Seventy-two hours after MPTP treatment, a drastic decrease in the intensity and number of immunopositive neurones and neurites in the SNpc (B), as well as in the intensity of TH immunolabelling in the whole striatum, is observed, especially in the dorsolateral area. (C,F) Two weeks after MPTP injection, a partial recovery of the immunolabelling is observed in both areas (C, SNpc; F, striatum). (G,H) Quantitative analysis confirms that changes observed 72 h and 2 weeks after MPTP injection, with respect to the control (ctrl), in both the number of TH+ cells/mm2 in the SNpc (G) and the optical density of the TH+ fibres in the striatum (H) are statistically significant. A partial, but significant, recovery is observed 2 weeks after the MPTP treatment in both SNpc and striatum. n = 4–5 animals/time point; bars represent the mean ± SEM; *P < 0.05, calculated by one-way ANOVA and Duncan test.

Figure S2. Persistent TH depletion induced by MPTP chronic treatment in monkeys. (A,B) TH-DAB immunolabelling in the SNpc (A1,2) and striatum (B1,2), both caudate (Cd) and putamen (Put), of control (A1,B1) and parkinsonian (A2,B2) monkeys. A marked decrease in TH&plus; cells and neurites in the SNpc (circled area in the left-hand drawing in A) and in the dopaminergic projections to the striatum (circled area in the left-hand drawing in B) is observed in parkinsonian macaques 2 years after the last MPTP injection, compared with control. (C) Quantification of the number of TH&plus; cells in the SNpc and measurement of the optical densities (O.D.) of the TH&plus;-immunopositive fibres in the striatum show that these decreases are statistically significant. Control group: n = 3 monkeys; Parkinsonian group: n = 5 monkeys. Histograms represent the mean ± SEM; *P < 0.05, **P < 0.01 calculated by the Student's t-test.

Figure S3. CNPase and MBP co-labelling in the mouse nigrostriatal pathway. Top row of confocal images shows a mouse striatum immunostained for CNPase (red) and MBP (green). An intense staining for both proteins decorates cell bodies (high magnifications are shown in the top row inserts, together with the lateral view along the z axis) and fibres coming from the cortex and crossing the striatum. Fibres of the nigrostriatal pathway also co-label for CNPase and MBP (bottom row magnifications). DAPI staining (blue) was used to label nuclei. Scale bars: 50 μm for the top-row pictures, 5 μm for the inserts and 10 μm for the bottom-row pictures.

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