Figure 2

Effects of exposure to embryonic ethanol (0.5% v/v, 22–24 hpf) and injection of cxcl12a mRNA (50 ng/µL) on the number of Hcrt neurons in the aAH, POA and pAH of 6 dpf Hcrt:EGFP zebrafish. (A) Bar graphs (n = 4–5/group) show the number of Hcrt neurons in the aAH, POA and pAH (Two-way ANOVA, Condition main effect: (F (2, 33) = 8.577, p = 0.0009), Brain area main effect: (F (2, 33) = 68.52, p < 0.0001), Condition ×Brain area interaction: (F (4, 33) = 3.431, p = 0.019), followed by Holm–Sidak post-hoc analysis). (B) Photomicrographs (25×, dorsal view) illustrate Hcrt neurons (green) that are located in the aAH, POA and pAH of Control + Vehicle (left), Ethanol + Vehicle (middle) and Control + cxcl12a mRNA (right) zebrafish. Scale bar: 10 µm. All results are shown as means ± standard errors. * p < 0.05, *** p < 0.001. Abbreviations: aAH: anterior part of the anterior hypothalamus, POA: preoptic area, pAH: posterior part of the anterior hypothalamus, Hcrt: hypocretin, hpf: hours post fertilization, dpf: days post fertilization.