Zebrafish embryos/larvae for rapid determination of effects on hypothalamic-pituitary-thyroid (HPT) and hypothalamic-pituitary-interrenal (HPI) axis: mRNA expression

To identify and prioritize chemicals that may affect thyroid and adrenal/interregnal endocrine system and to reduce cost and animal use by conventional toxicity assay, an in vivo screening assay was developed using zebrafish embryos/larvae based on measurement of expression of genes that were suggested to play important roles in hypothalamic-pituitary-thyroid (HPT) and hypothalamic-pituitary-interrenal (HPI) axis. Model chemicals that could modulate HPT and HPI axis in adult fish were selected in assay validation, including anti-thyroid agent 6-Propyl-2-thiouracil (PTU) and cytochrome P450 11B (Cyp11b) enzyme inhibitor metyrapone (MET). Zebrafish embryos were exposed to different concentrations of model chemical from 4 h post-fertilization (hpf) to 5 d post-fertilization (dpf). Exposure to PTU increased mRNA expression of sodium iodide symporter (nis) and thyroglobulin (tg) involved in HPT axis, and MET treatment up-regulated all the mRNA expression tested involved in HPI axis by a compensatory mechanism. These results suggested that HPT and HPI axis were active upon chemical exposure at least at 5 dpf zebrafish. Furthermore, we studied the effects of PTU or MET on the cross-talk between HPT and HPI axis. The results demonstrated that PTU and MET could affect cross-talk responses in zebrafish embryos/larvae.