Table III. Body length (in mm) at 30 hpf in ethanol and retinoic acid treated embryos.
Embryos were treated from 3 hpf until 24 hpf (during gastrulation and somitogenesis) with 100 mM ethanol, 10−9 M retinoic acid, or 100 mM ethanol and 10−9 M retinoic acid together, and compared to untreated control embryos. At 30 hpf embryos were fixed using paraformaldehyde to stop development, and these embryos were imaged using a stereo microscope. Embryo length was measured along the dorsal surface of the embryos from the most anterior point where the embryo proper touches the yolk to the tip of the tail using ImageJ image analysis software (see Materials and Methods). Two-way ANOVA was performed to compare treatment groups. Ethanol-by-retinoic acid interaction was significant (P=0.0005), and pairwise comparison P-values are shown. Bonferroni adjustment for multiple (six) comparisons was used to ensure an overall ninety five percent confidence. Thus, P-value≤0.008 was used to conclude that groups were statistically different.
| Control | Ethanol | Ethanol+RA | RA only | |
|---|---|---|---|---|
| Mean | 3.39 | 3.10 | 3.23 | 3.26 |
| SD | 0.12 | 0.25 | 0.12 | 0.12 |
| n | 19 | 19 | 21 | 19 |
| P (vs Control) | <0.0001 | 0.0026 | 0.0206 | |
| P (vs Ethanol) | 0.0068 | 0.0011 | ||
| P (vs RA) | 0.4895 |