Figure 5:

Gad1 gene expression is reduced in 24 hpf embryos following acute 1% ethanol and 3 mg/L ACEA co-exposure from 8–10 hpf and is rescued by fgf8 mRNA. Embryos were analyzed at 24 hpf for gad1 gene expression by whole-mount in situ hybridization. A, wild-type embryos; B, 0.5% ethanol alone; C, 1 mg/L ACEA alone; D, embryos injected with fgf8 mRNA alone at one- to two-cell stage; E, embryos exposed to 1% ethanol alone; F, embryos exposed to 3mg/L ACEA alone; G, embryos co-exposed to 0.5% ethanol and 3 mg/L ACEA; H, embryos co-exposed to 3 mg/L ACEA and 1% ethanol; I, embryos injected with fgf8 mRNA at one- to two-cell stage and co-exposed to 3 mg/L ACEA and 1% ethanol. Note: Decreased gad1 gene expression was only observed with combined 1% ethanol and 3 mg/L ACEA treatment, and fgf8 mRNA overexpression was able to reverse this reduced gene expression (F). N≥ 30/group.