Figure 5

Th/th+ neurons reside in the tegmental midbrain and the rostral hindbrain. (A–F”, F1–F3) Whole-brain fluorescent in situ hybridization and immunofluorescence staining for the indicated mRNAs (magenta) and GFP (green) in th^{m1512Tg(2A−QF2)}; Tg(QUASr:GFP)c403 larval brains at 120 hpf. Dorsal views of z-projections (B') or single planes (A, B, C–F”, F1-F3). Anterior is to the left. (A, B) Expression of the midbrain–hindbrain boundary marker pax2a in comparison with GFP in a dorsal view (A) and a more ventral view (B). The dashed box indicates GFP+ cells in the midbrain. (B') z-projection (z-stack size: 31 μm) and magnification of the cells marked by a dashed box in (B). (C) Expression of the midbrain–hindbrain boundary marker en1a/en2a in comparison with GFP. (D, E) Expression of the mammalian midbrain dopaminergic marker pitx3 in comparison with GFP, focusing on GFP+ cells in the midbrain (D) and a more ventral view (E). (F–F”) Expression of the mammalian mDA and tegmental marker nr4a2a/nr4a2b in comparison with GFP. The dashed box in (F”) indicates GFP+ cells in the midbrain. (F1–F3) Magnification of the cells marked by a dashed box in (F”). Scale bars: [(A), also for (B, C–F”)] 100 μm; (B') 20 μm; (F1–F3) 10 μm. For better representation of low and high signal intensities, non-linear adjustments were made to whole image panels (see Section 2.8). LC, locus coeruleus; Mb, midbrain; MHB, midbrain–hindbrain boundary; Pr, pretectum; rHb, rostral hindbrain; Teg, tegmentum.