Aryl hydrocarbon receptor 2 mediates 2,3,7,8-tetrachlorodibenzo-p-dioxin developmental toxicity in zebrafish
Aryl hydrocarbon receptor 2 mediates 2,3,7,8-tetrachlorodibenzo-p-dioxin developmental toxicity in zebrafish
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DOI：
10.1093/toxsci/kfg202
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发表时间：
2003-11-01
影响因子：
3.8
通讯作者：
Peterson, RE
中科院分区：
文献类型：
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作者：
Prasch, AL;Teraoka, H;Peterson, RE
为了使用斑马鱼作为模型脊椎动物研究 2,3,7,8-四氯二苯并-对二恶英 (TCDD) 的发育毒性，有必要了解斑马鱼芳烃受体 (AHR) zfAHR1 或 zfAHR2 中的一种或两种形式是否介导毒性。为了确定 zfAHR2 的作用，使用反义吗啉方法来抑制该蛋白质的翻译。在未经 TCDD 处理的胚胎中，未发现 zfahr2 吗啉代 (zfahr2-MO) 对正常发育有影响。在1-2细胞阶段的胚胎中注射zfahr2-MO可减少TCDD诱导的zfCYP1A mRNA转录，直至受精后96小时（hpf），并且在72 hpf时对胚胎中zfCYP1A蛋白的免疫组织化学检测显示表达显着降低。 zfahr2-MO完全保护胚胎免受TCDD引起的水肿和贫血，并提供针对TCDD引起的外周血流量减少的保护。然而，当吗啉不再有效时，后来观察到血流量略有减少。由于 TCDD 的持续存在以及 zfahr2-MO 的有效性随着时间的推移而降低，吗啉代仅提供短暂的保护，以防止 TCDD 诱导的下颌软骨形成抑制，并且不能防止 TCDD 在发育后期启动的效应，即阻碍鱼鳔膨胀。 zfahr2-MO 并不能保护胚胎免受 TCDD 诱导的死亡，但确实使其发病延迟了 48 小时。 zfahr2 morphant在发育后期（超过144 hpf）中表现出的TCDD发育毒性终点与注射对照吗啉代的暴露于TCDD的胚胎明显不同。最引人注目的是，暴露于 TCDD 的 zfahr2 突变体从未出现水肿。综上所述，这些结果表明 zfAHR2 介导斑马鱼 TCDD 发育毒性的几个终点。
In order to use the zebrafish as a model vertebrate to investigate the developmental toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), it is essential to know whether one or both forms of the zebrafish aryl hydrocarbon receptor (AHR), zfAHR1 or zfAHR2, mediate toxicity. To determine the role of zfAHR2, an antisense morpholino approach was used to knock down translation of the protein. No effect of the zfahr2 morpholino (zfahr2-MO) was seen on normal development in embryos not treated with TCDD. Injection of embryos at the 1-2 cell stage with zfahr2-MO decreased TCDD-induced transcription of zfCYP1A mRNA until 96 h post fertilization (hpf), and immuno-histochemical detection of zfCYP1A protein in embryos at 72 hpf revealed a dramatic decrease in expression. The zfahr2-MO completely protected embryos from TCDD-induced edema and anemia and provided protection against TCDD-induced reductions in peripheral blood flow initially; however, a slight reduction in blood flow was observed at later times when the morpholino was no longer effective. Due to persistence of TCDD and decreasing effectiveness of the zfahr2-MO over time, the morpholino provided only transient protection against TCDD-induced inhibition of chondrogenesis of the lower jaw, and no protection against an effect of TCDD that was initiated late in development, blockade of swimbladder inflation. The zfahr2-MO did not protect embryos from TCDD-induced mortality but did produce a 48 h delay in its onset. Endpoints of TCDD developmental toxicity manifested in zfahr2 morphants at late stages of development, beyond 144 hpf, were clearly different from TCDD-exposed embryos injected with a control morpholino. Most strikingly, zfahr2 morphants exposed to TCDD never developed edema. Taken together, these results demonstrate that zfAHR2 mediates several endpoints of TCDD developmental toxicity in zebrafish.