An ex vivo assay for screening glucocorticoid signaling disruption based on glucocorticoid-response gene transcription in Xenopus tails

There is a pressing need for developing in vivo or ex vivo assays to screen the glucocorticoid (GC) signaling disruption of chemicals. Thus, we aimed to establish an ex vivo assay for screening GC signaling disruption based on the GC-response gene transcription in Xenopus laevis tails cultured ex vivo. Firstly, we investigated effects of corticosterone (CORT, a main GC in frogs) on GC-response gene expression, and determined the six genes as molecular endpoints for assaying the GC signaling disruption. CORT in the range of 1.56-400nmol/L was found to up-regulate transcription of the six GC-response genes, exhibiting comparable or higher sensitivity than previously reported assays. To validate this ex vivo assay, then, we examined effects of dexamethasone (a known GC signaling agonist) on GC-response gene expression. Dexamethasone displayed an agonistic action in a concentration-dependent manner, further demonstrating the efficiency of the established assay. Finally, we applied the ex vivo assay to evaluate the GC signaling disruption of bisphenol A (BPA). In accordance with previous reports, we found a concentration-dependent agonistic activity of BPA, showing that the established assay is effective for detecting the GC signaling disrupting activity of environmental chemicals. Correspondingly, the GC signaling agonistic actions of CORT and BPA in ex vivo tails accorded with the observations in vivo, indicating that the ex vivo assay is able to detect the actions of chemicals in vivo. Overall, we established an ex vivo assay that can effectively screen GC signaling disruption of environmental chemicals.