Graduate School of Agriculture, Ehime University, 3-5-7 Tarumi, Matsuyama, 790-8566, Japan.
Department of Creative Engineering, National Institute of Technology, Ariake College, 150 Higashi-Hagio, Omuta, Fukuoka, 836-8585, Japan.
Ecotoxicol Environ Saf. 2020 Apr 15;193:110324. doi: 10.1016/j.ecoenv.2020.110324. Epub 2020 Feb 20.
This study assessed the transcription levels of estrogen-responsive genes, such as vitellogenins (Vtg1 and Vtg2), choriogenins (ChgL, ChgH, and ChgHm), cytochrome P450 aromatase (CYP19a1b), and ER subtypes (ERα, ERβ1, and ERβ2), in 7 days-post-fertilization (dpf) embryos and 9 and 12 dpf larvae of medaka (Oryzias latipes) exposed to estrogenic endocrine-disrupting chemicals (EDCs). The <5 h-post-fertilization embryos were exposed to EDCs such as 17β-estradiol (E2), p-n-nonylphenol (NP), and bisphenol A (BPA). In E2 (0.10-222 nM)-treated 7 dpf embryos and 9 or 12 dpf larvae, ChgL, ChgH, and ChgHm expression was up-regulated in a concentration-dependent manner. By contrast, interestingly, Vtg1 and Vtg2 expression was not induced in E2-treated 7 dpf embryos but was significantly induced in 9 and 12 dpf larvae, suggesting a developmental-stage-specific regulatory mechanism underlying Vtg expression. The maximum concentrations of NP (0.09-1.5 μM) and BPA (1.8-30 μM) up-regulated Chg expression in 9 or 12 dpf larvae, and the relative estrogenic potencies (REPs) of E2, NP, and BPA were 1, 2.1 × 10, and 1.0 × 10, respectively. Chg messenger RNA (mRNA) in medaka embryos and larvae can be used as a sensitive biomarker for screening potential estrogenic EDCs. Our assay system using embryos and larvae can be used as an in vivo alternative model because independent feeding stages (e.g., embryonic and early larval stages) are suitable alternatives.
本研究评估了卵黄蛋白原(Vtg1 和 Vtg2)、绒毛蛋白(ChgL、ChgH 和 ChgHm)、细胞色素 P450 芳香酶(CYP19a1b)和雌激素受体亚型(ERα、ERβ1 和 ERβ2)等雌激素反应基因在受精后 7 天(dpf)胚胎和 9 天和 12 天的黄颡鱼(Oryzias latipes)幼虫中的转录水平,这些幼虫接触了雌激素类内分泌干扰化学物质(EDCs)。受精后 <5 小时的胚胎暴露于 EDC 如 17β-雌二醇(E2)、对壬基苯酚(NP)和双酚 A(BPA)。在 E2(0.10-222 nM)处理的 7 dpf 胚胎和 9 或 12 dpf 幼虫中,ChgL、ChgH 和 ChgHm 的表达呈浓度依赖性上调。相比之下,有趣的是,E2 处理的 7 dpf 胚胎中未诱导 Vtg1 和 Vtg2 的表达,但在 9 和 12 dpf 幼虫中显著诱导,表明 Vtg 表达存在发育阶段特异性调节机制。NP(0.09-1.5 μM)和 BPA(1.8-30 μM)的最大浓度上调了 9 或 12 dpf 幼虫的 Chg 表达,E2、NP 和 BPA 的相对雌激素效力(REP)分别为 1、2.1×10 和 1.0×10。黄颡鱼胚胎和幼虫的 Chg 信使 RNA(mRNA)可作为筛选潜在雌激素 EDC 的敏感生物标志物。我们使用胚胎和幼虫的检测系统可以作为一种体内替代模型,因为独立的喂养阶段(如胚胎和早期幼虫阶段)是合适的替代物。