| This project sought to explore the role of the JNK signaling pathway and c-Jun in cadmium-induced embryotoxicity and their role in normal embryo development. Primary cultures of mouse embryo limb bud cells harvested at day 11 of gestation were pre-treated with JNK inhibitor SP600125 and antisense oligonucleotide (ASO) c-Jun, and then incubated with or without cadmium chloride. Various endpoints of toxicity were measured through cell proliferation and viability assays, cytotoxicity assays (Neutral Red), and measurement of differentiation (Alcian Blue Assay). It was found that cadmium-induced activation of JNK and c-Jun were inhibited by SP600125 in a dose-dependent manner. However, cadmium-induced cytotoxicity could not be prevented by SP600125. Interestingly, our results showed that cadmium-induced c-Jun activation and embryotoxicity could both be partially prevented by ASO c-Jun, but that ASO c-Jun had no effect on JNK activation. In addition, our observations also demonstrated that inhibiting basal activation of JNK and c-Jun can decrease limb bud cells proliferation, survival and differentiation during normal growth condition. (Abstract shortened by UMI.)... |
