Fig. S2

Fig. S2 (related to Fig. 2) Dopaminergics influence development of Islet- 1:GFP+ motor neurons, but not Pax2a+ or Glyt2:GFP+ interneurons. A: Lateral views of the trunk region of living islet-1:GFP transgenic animals under a stereo-microscope are shown. The dopamine agonist NPA (drug exposure 24 à 51 hpf; ANOVA, ***P < 0.001 compared to DMSO) significantly increases and the antagonist Haloperidol (drug exposure 24 to 56 hpf; ANOVA, **P < 0.01 compared to DMSO) decreases the percentage of embryos with islet- 1:GFP+ motor neurons (arrows) in a dose dependent fashion. B: The D4 specific antagonist L-745870 attenuates development of islet-1:GFP+ motor neurons, which cannot be overcome by the addition of NPA (56 hpf; ANOVA, ***P < 0.0001). C: Visual scoring of islet-1:GFP embryos is confirmed by cell counts and fluorescence intensity measurements. Lateral views of z-projected confocal image stacks of the mid trunk region are shown. At 51 hpf (left column), cell number (*P < 0.05; Mann-Whitney U-test, one tailed) and fluorescence intensity are significantly increased (*P < 0.05; Mann-Whitney Utest, one tailed) in NPA treated embryos. At 56 hpf (right column), cell number (ANOVA, ***P < 0.0001) and fluorescence intensity (ANOVA, ***P < 0.0001) are significantly lower in L-745870 treated embryos and still increased in NPA treated animals compared to DMSO controls (ANOVA, *P < 0.05; **P < 0.01). Fluorescence intensity of an expression domain in the cerebellum was unchanged by any of the treatments (data not shown). D: Lateral trunk views of embryos at 33 hpf are shown (drug exposure 24 to 33 hpf; genotypes and drugs are indicated). NPA has no effect on the numbers of glyt2:GFP+ or pax2a:GFP+ interneurons. Error bars represent SEM. Scale bars: A: 100 μm; C: 60 μm, D: 50 μm.