DEV002329 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Localisation of major DA cell groups in the newt brain. (A,B) Olfactory bulb. (C) DA fibres within the striatum (Str). (D) Preoptic area (POa). (E,F) Suprachiasmatic nucleus (SCN) and nucleus of the ventral thalamus (VTn). (G) Tuberculum posterius medialis/lateralis (TPm/TPl). (H) Rostral tegmental area of the midbrain, TM. (I) Caudal TM. (J) Hindbrain. (E′-H′) High-power micrographs of E-H, showing the DA populations counted, which were consistently lesioned by 6-OHDA administration. Scale bars: 400 μm in A-J; 100 μm in E′-H′.

• Supplemental Figure S2 -

  Fig. S2. Regeneration of TH⁺ cells along the rostrocaudal axis of the diencephalon and mesencephalon. (A-E) sham; (F-J) 3 days post-lesion; (K-O) 30 days post-lesion. Scale bar: 1 mm.

• Supplemental Figure S3 -

  Fig. S3. Temporal regulation of engrailed 1 (En1) expression. (A) Number of En1⁺ cells at 3, 8 and 13 days after lesioning. (B) Proportion of En1⁺ to TH⁺ cells during DA regeneration. Mean±s.e.m; ANOVA with Tukey post-hoc test; **, P<0.005. Note that En1 expression preceded that of TH. (C) Example of TH (green) and En1 (red) double-positive cells in the TM of a control animal, illustrating the expression of En1 in all TH⁺ neurons.

• Supplemental Figure S4 -

  Fig. S4. Regulation of Msx1/2 expression in the dorsal diencephalon and mesencephalon after 6-OHDA injection. Note that in contrast to the ventral regions, there is no significant upregulation at 7 days. Mean±s.e.m; ANOVA with Tukey post-hoc test; *, P<0.05.

• Movie 1 -

  Movie 1. Movie capture of amphetamine-induced locomotor response. Typical sham-lesioned (lower panel) and 6-OHDA-lesioned (upper panel) animals following a single amphetamine challenge 3 days after surgery.

• Movie 2 -

  Movie 2. Movie capture of locomotor activity without amphetamine challenge. Typical sham-lesioned (upper panel) and 6-OHDA-lesioned (lower panel) animals without amphetamine administration 3 days after surgery.

• Movie 3 -

  Movie 3. Confocal analysis of TH BrdU double immunostaining. Confocal planes in three dimensions of two double-positive TH⁺(green) BrdU⁺ (red) cells in the lesioned brain.