Fig. 3

GFP expression in huORFZ as signals of cell under acute and chronic toxic stresses.

At 72huORFZ embryos were treated with Cadmium (Cd²⁺; A-I) or Copper (Cu²⁺; J-R). (A). The survival rate of huORFZ embryos incubated in Cd²⁺ with different concentrations and treatment time. Open bars indicate huORFZ embryos that remained in the Cd²⁺ solution throughout the experiment; black bars indicate the embryos removed for recovery in clean water after 24 hr of Cd²⁺ treatment. N/A indicates that prolonged treatment was not conducted as a result of 100% lethality. (B-E). GFP signals immediately or 24, 48 and 72 hr after 24 hr of Cd²⁺ treatment. (F-I). TUNEL assay at the olfactory epithelium immediately or 72 hr after 24 hr of Cd²⁺ treatment. Red color represents TUNEL assay; Green color represents GFP signal; and Blue color represents DAPI staining. (J). The survival rates of huORFZ embryos incubated in Cu²⁺ with different concentrations and treatment times. The experimental strategy was the same as A-I. (K-N). GFP signals immediately or 24, 48 and 72 hr after 24 hr of Cu²⁺ treatment. (O-R). TUNEL assay at the skin immediately or 72 hr after 24 hr of Cu²⁺ treatment. (S-U). For the chronic toxicity test, 72 hpf huORFZ embryos were treated with chlorpyrifos with a concentration (86 nM) below WHO guidelines. (S). The survival rates of the embryos treated with chlorpyrifos for zero to three days. (T, U). GFP signals immediately or 72 hr after three days of chlorpyrifos treatment. (F-I). are transverse section with dorsal to the top. All other images are lateral views with anterior to the left. For each treatment (each bar in A, J and S), n = 100 embryos evenly distributed in five repetitions.