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Fig. 4 The effects of disrupted dopaminergic signaling during development on neural circuit formation. A, C, and E: Tubulin immunostaining in the control group, dopamine antagonist–treated group (0.2 μmol/L SCH 23390), and otp mutant zebrafish at 48 hours after fertilization. The lines indicate the measurement of anterior commissure (AC) and post-optic commissure (POC) in the midline, respectively. B, D, and F: SV2 immunostaining in the control group, dopamine antagonist–treated group (0.2 μmol/L SCH 23390), and otp mutant zebrafish at 48 hours after fertilization. The embryo was oriented in rostral-caudal axis, and the rostral part was on top. The irregular shape along the outer edge of synaptic vesicle glycoprotein 2A (SV2) staining was used to measure the area in the AC or POC region. G and H: Quantification of AC and POC width stained by tubulin. Significant differences in AC [analysis of variance, F(2,15) = 5.058] and POC widths [analysis of variance, F(2,15) = 10.54], stained with tubulin, were observed between the control and SCH 23390–treated groups (df = 15) and the control and otp knockout (KO) groups (df = 15) as determined by one-way analysis of variance followed by the Dunnett test. I and J: Quantification of area in the AC [analysis of variance, F(2,14) = 13.85] and POC regions [analysis of variance, F(2,14) = 3.651; P = 0.053] stained by SV2. Significant differences were observed between the control and SCH 23390–treated or KO otp groups using one-way analysis of variance followed by the Dunnett multiple comparisons test. No significant differences were found in the normalized area of the POC among the three groups. ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001. Scale bar = 50 μm. Ctrl, control.