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Re-evaluation of thyroid hormone signaling antagonism of tetrabromobisphenol A for validating the t3-induced Xenopus metamorphosis assay

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Journal of Environmental Sciences 2017年第2期29卷 325-332页

作者： Yao Wang Yuanyuan Li Zhanfen Qin Wuji Wei College of Environment Nanjing University of TechnologyNanjing 211816China State Key Laboratory of Environmental Chemistry and Ecotoxicology Research Center for Eco-environmental SciencesChinese Academy of SciencesBeijing I0008SChina University of Chinese Academy of Sciences Beijing 100049China

We developed the t3-induced Xenopus metamorphosis assay, which is supposed to be able to sensitively detect thyroid hormone（tH） signaling disruption of chemicals. the present study aimed to validate the t3-induced Xenopus metamorphosis assay by re-evaluating the tH signaling antagonism of tetrabromobisphenol A（tBBPA）, a known tH signaling disruptor. According to the assay we developed, Xenopus tadpoles at stage 52 were exposed to 10–500 nmol/L tBBPA in the presence of 1 nmol/L t3. After 96 hr of exposure, tBBPA in the range of 10–500 nmol/L was found to significantly inhibit t3-induced morphological changes of Xenopus tadpoles in a concentration-dependent manner in term of body weight and four morphological endpoints including head area（HA）, mouth width（MW）, unilateral brain width/brain length（ULBW/BL）, and hind-limb length/snout-vent length（HLL/SVL）.the results show that these endpoints we developed are sensitive for characterizing the antagonistic effects of tBBPA on t3-induced metamorphosis. Following a 24-hr exposure,we found that tBBPA antagonized expression of t3-induced tH-response genes in the tail,which is consistent with previous findings in the intestine. We propose that the tail can be used as an alternative tissue to the intestine for examining molecular endpoints for evaluating tH signaling disruption. In conclusion, our results demonstrate that the t3-induced Xenopus metamorphosis assay we developed is an ideal in vivo assay for detecting tH signaling disruption.

关键词： thyroid hormone signaling tetrabromobisphenol A Xenopus metamorphosis t3 induction tail

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Optimization of the t3-induced Xenopus metamorphosis assay for detecting thyroid hormone signaling disruption of chemicals

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Journal of Environmental Sciences 2017年第2期29卷 314-324页

作者： Xiaofang Yao Xiaoying Chen Yinfeng Zhang Yuanyuan Li Yao Wang Zongming Zheng Zhanfen Qin Qingdong Zhang School of Environment and Resource Southwest University of Science and TechnologyMianyang 621010China State Key Laboratory of Environmental Chemistry and Ecotoxicology Research Center for Eco-environmental SciencesChinese Academy of SciencesBeijing 100085China University of Chinese Academy of Sciences Beijing 100049China

t3-induced Xenopus metamorphosis is an ideal model for detecting thyroid hormone（tH）signaling disruption of chemicals. to optimize the t3-induced Xenopus assay and improve its sensitivity and reproducibility, we intend to develop quantitatively morphological endpoints and choose appropriate concentrations and exposure durations for t3 *** laevis at stage 52 were exposed to series of concentrations of t3（0.31–2.5 nmol/L）for 6 days. By comparing morphological changes induced by t3, we propose head area,mouth width, unilateral brain width/brain length, and hindlimb length/snout-vent length as quantitative parameters for characterizing t3-induced morphological changes, with body weight as a parameter for indicating integrated changes. By analyzing time-response curves, we found that following 4-day exposure, t3-induced grossly morphological changes displayed linear concentration–response curves, with moderate morphological changes resulting from 1.25 nmol/L t3 exposure. When using grossly morphological endpoints to detect tH signaling disruption, we propose 4 days as exposure duration of t3, with concentrations close to 1.25 nmol/L as induction concentrations. However, it is appropriate to examine morphological and molecular changes of the intestine on day 2 due to their early response to t3. the quantitative endpoints and t3 induction concentrations and durations we determined would improve the sensitivity and the reproducibility of the t3-induced Xenopus metamorphosis assay.

关键词： thyroid hormone signaling Disruption Xenopus laevis t3 induction Morphological endpoints tH-response gene expression

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