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Developing a novel quantitative parameter for characterizing spatial distribution of fish following exposure to chemicals and wastewater:Behavioral Gini coefficient

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Journal of Environmental Sciences 2024年第7期141卷 129-138页

作者： Xuanyue Chen Yuanyuan Li zhanfen qin 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

While the spatial distribution pattern of fish is increasingly used for toxicological test o chemicals or wastewater,no ideal parameter is available for quantitative assessment of spa tial distribution,especially uneven distribution with multiple ***,to develop a quantitative assessment parameter for spatial distribution,the zebrafish were exposed to ethanol,pentylenetetrazole(PTZ),paraquat dichloride(paraquat)and wastewater,followed by a behavioral test in a narrow *** data was acquired and analyzed by id Tracker and *** comparing the effects of all treatments on behavior parameters we confirmed that the spatial distribution was more easily altered rather than general loco motor parameters,e.g.0.7-70 mg/L PTZ and 5-20 mg/L paraquat being effective for altering spatial distribution but having little effects on general locomotor *** on the heatmap,i.e.,the cumulative proportion of grids and that of frequency in grids,we calcu lated the behavioral Gini coefficient(G_(b))for quantitative assessment of fish spatial distri *** Gini coefficient ranged from zero to 1,with larger values meaning poorer even ness of spatial *** note,G_(b)showed smaller coefficient of variations(CV)with3%-19%between replicate tanks in all treatments than the highest frequency(4%-79%),dis playing well ***,G_(b)addressed the challenge of the complicated heatmap with multiple ***,the behavioral Gini coefficient we established is an idea parameter to quantitatively assess spatial distribution of fish shoal,which is expected to be applied in toxicity testing for chemicals and wastewater and automatic quality monitoring for surface water and aquaculture water.

关键词： Zebrafish Behavioral test Spatial distribution Behavioral Gini coefficient

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Transcriptomic analysis reveals the effects of maternal exposure to bisphenol AF on hypothalamic development in male neonatal mice

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Journal of Environmental Sciences 2024年第7期141卷 304-313页

作者： Lin Lv Yuanyuan Li Xuanyue Chen zhanfen qin 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

Fragmented data suggest that bisphenol AF(BPAF),a chemical widely used in a variety of products,might have potential impacts on the ***,we employed male neonatal mice following maternal exposure to explore the effects of low-dose BPAF on hypothalamic development by *** found that maternal exposure to approximately 50μg/(kg·day)BPAF from postanal day(PND)0 to PND 15 altered the hypothalamic transcriptome,primarily involving the pathways and genes associated with extracellular matrix(ECM)and intercellular adhesion,neuroendocrine regulation,and neurological *** RNA analysis confirmed the changes in the expression levels of concerned ***,we further revealed that low-dose BPAF posed a stimulatory impact on pro-opiomelanocortin(POMC)neurons in the arcuate nucleus of the hypothalamus and induced the browning of inguinal white adipose *** findings indicate that developmental exposure to low-dose BPAF could interfere with hypothalamic development and thereby lead to alterations in the ***,5000μg/(kg·day)BPAF caused slighter,non-significant or even inverse alterations than the low dose of 50μg/(kg·day),displaying a dose-independent *** observations suggest that the the dose-independent effects of BPAF might be associated with oxidative stress and inflammatory responses caused by the high ***,our study highlights a risk of low-dose BPAF to human neuroendocrine regulation and metabolism.

关键词： Bisphenol AF Hypothalamus Transcriptomic alteration Pro-opiomelanocortin(POMC)neurons White adipose tissue browning

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Tetrabromobisphenol A exerts thyroid disrupting effects but has little overt impact on postnatal brain development and neurobehaviors in mice

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Journal of Environmental Sciences 2024年第8期142卷 1-10页

作者： Shilin Song Yuanyuan Li Lin Lv Mengqi Dong zhanfen qin State Key Laboratory of Environmental Chemistry and Ecotoxicology Research Center for Eco-Environmental SciencesChinese Academy of SciencesBeijing 100085China College of Resources and Environment University of Chinese Academy of SciencesBeijing 100049China School of Environment Tsinghua UniversityBeijing 100084China

Tetrabromobisphenol A(TBBPA)is a widely used brominated flame *** is evidence showing that TBBPA can exert thyroid disrupting effects in mammals,but different results were also reported,along with inconsistent reports regarding its ***,we investigated thyroid disrupting effects and neurotoxicity of TBBPA(5,50,500μg/(kg·day))to male mice following maternal and direct exposure through drinking water,with the antithyroid drug propylthiouracil(PTU)as the positive *** postnatal day(PND)15,we expectedly observed severe thyroid compensatory hyperplasia and cerebellar developmental retardation in PTU-treated *** highest dose of TBBPA also caused thyroid histological alteration but had no effects on cerebellar development in terms of Purkinje cell morphology and the thickness of the internal granular layer and the molecular layer of the *** puberty and adulthood,the thyroid morphological alterations became more pronounced in the TBBPA-treated animals,accompanied by decreased serum thyroid hormone ***,the 50 and 500μg/(kg·day)TBBPA groups showed a significant decrease in the serum level of serotonin,a neurotransmitter associated with anxiety ***,the highest dose group displayed anxiety-like behaviors in the elevated plus-maze test on PND 35,but this neurobehavioral alteration disappeared on PND ***,no changes in neurobehavioral parameters tested were found in TBBPAtreated animals at puberty and ***,all observations show that TBBPA can exert thyroid disrupting effects but has little overt impact on brain development and neurobehaviors in mice,suggesting that thyroid disruption does not necessarily cause overtly adverse neurodevelopmental outcomes.

关键词： Tetrabromobisphenol A Thyroid disruption Brain development Neurobehavior Cerebellum

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基于两栖动物的甲状腺干扰物测试方法分析

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生态毒理学报 2024年第3期19卷 131-139页

作者：彭九妹李圆圆秦占芬何益得张永军南京工业大学环境科学与工程学院南京211816 中国科学院生态环境研究中心北京100085 中国科学院大学北京100049

两栖动物经历一个主要由甲状腺激素(TH)调控的变态发育过程,表现为外部形态和内部组织器官乃至TH靶基因表达的剧烈变化,这为研究和测试甲状腺干扰物提供了一个良好的模型。自2009年以来,经济合作组织(OECD)相继发布了3个基于两栖动物的... 详细信息

两栖动物经历一个主要由甲状腺激素(TH)调控的变态发育过程,表现为外部形态和内部组织器官乃至TH靶基因表达的剧烈变化,这为研究和测试甲状腺干扰物提供了一个良好的模型。自2009年以来,经济合作组织(OECD)相继发布了3个基于两栖动物的甲状腺干扰物测试方法导则,即两栖动物变态试验(amphibian metamorphosis assay,AMA)导则TG 231、两栖动物幼体生长和发育试验(larval amphibian growth and development assay,LAGDA)导则TG 241和爪蛙活动胚胎甲状腺试验(X enopus eleutheroembryo thyroid assay,XETA)导则TG 248。目前这些方法已被纳入OECD的内分泌干扰物测试和评估概念框架,用于筛查测试甲状腺干扰物。不同于以自然变态发育为基础的AMA和LAGDA,国内学者发展了筛查甲状腺干扰物的T3诱导爪蛙变态试验(T3-induced Xenopus metamorphosis assay,TiXMA)。为促进对这些方法的理解和应用,本文综述了各方法的原理、终点指标、结果解释和应用现状,初步比较了各方法测试甲状腺干扰物的敏感性和特异性,并提出通过开展更多化学物质测试以对方法进行验证的必要性。

关键词：甲状腺干扰物两栖动物变态发育测试方法

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Application of Xenopus laevis in ecotoxicology （Ⅰ）--Introduction and quality control of laboratory animal

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Chinese Science Bulletin 2006年第11期51卷 1273-1280页

作者： qin zhanfen XU Xiaobai State Key Laboratory of Environmental Chemistry and Ecotoxicology Research Center for Eco-environmental Sciences Chinese Academy ofSciences Beijing 100085 China

The aim of the series of papers is to discuss the application of Xenopus laevis, as model animal in biology, in ecotoxicology. X. laevis as model animal is wildly used in biological study and has pro- vided a lot of relating data because of many advan- tages, such as living in water and being easily main- tained, laying eggs in the whole year, and externally fertilizing and developing. Embryos and larvae of X. laevis like other amphibians are directly exposed in the aquatic environment and sensitive to pollutants. In addition, sex differentiation and sex organ devel- opment of X. laevis are sensitive to sex hormones and endocrine disruptors with sex hormone activities, which enable X. laevis to be used in studies on sex hormone disruption and reproductive toxicity of en- docrine disruptors. Metamorphic development of X. laevis is very sensitive to thyroid hormones and thy- roid disruptors, which enables X. laevis to be used for evaluating thyroid disruptors. Also, X. laevis ecotoxi- cology can be linked with amphibian population de- clines and malformed frog occurrence, being one of the hotspots in ecology. Thus, more and more labo- ratories have introduced X. laevis to ecotoxicological study. The quality of laboratory animals correlates with scientificity and reliability of results from animal experiments. It is especially important for toxicology. Quality control of X. laevis involving several factors such as water and food is discussed in this paper.

关键词：有爪蟾蜍生态毒理学实验动物质量控制

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

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Journal of Environmental Sciences 2018年第4期30卷 104-112页

作者： Xiaoying Chen Yuanyuan Li Min Zhu zhanfen qin State Key Laboratory of Environmental Chemistry and Ecotoxicology Research Center for Eco-Environmental Sciences Chinese Academy of Sciences Beijing 100085 China University of Chinese Academy of Sciences Beijing 100049 China School of Environmental Sciences and Engineering Nanjing Tech University Nanjing 211816 China

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 1.56–400 nmol/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.

关键词： Glucocorticoid signaling Glucocorticoid-response gene Xenopus laevis tail Dexamethasone Bisphenol A(BPA)

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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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Effects of triclosan on gonadal differentiation and development in the frog Pelophylax nigromaculatus

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Journal of Environmental Sciences 2018年第2期30卷 157-165页

作者： Juan Chen Tan Meng Yuanyuan Li Kun Gao zhanfen qin State Key Laboratory of Environmental Chemistry and Ecotoxicology Research Center for Eco-Environmental Sciences Chinese Academy of Sciences Beijing 100085 China University of Chinese Academy of Sciences Beijing 100049 China

Previous studies have reported that triclosan（TCS） could possess an androgenic *** aimed to investigate the effects of TCS on gonadal differentiation and development in the frog Pelophylax nigromaculatus, a sensitive amphibian species to androgenic chemicals.P. nigromaculatus tadpoles at stage 24 were exposed to TCS（3, 30, and 300 nmol/L） to stage46 in a semi-static exposure system. At the end of exposure, gonadal morphology and histology, sex ratio and gonadal expression of sex-biased genes were examined in P. nigromaculatus. In each TCS treatment group, we found several individuals whose gonads exhibited morphological and/or histological abnormalities. Gonadal histological abnormalities were characterized by few oocytes and many somatic cells. Although the percentage of the individuals with abnormal gonads was low（7.8%） among all animals treated with TCS,statistical test revealed the sex ratios in the 3 and 300 nmol/L TCS treatment groups were significantly different from the solvent control. In the 30 nmol/L TCS treatment group,abnormal gonads were also observed, although the sex ratio was not changed compared with the solvent control, which was possibly due to the smaller sample size in this group. In all the TCS treatment groups, the sex ratios were not obviously male-biased, but the expression levels of some sex-biased genes were significantly altered by TCS. Altogether,our results suggest that TCS, even at environmentally relevant concentrations, could disrupt gonadal differentiation and development in P. nigromaculatus, but we are not sure whether the disrupting effects were associated with masculinization or feminization.

关键词： Triclosan Pelophylax nigromaculatus Gonadal abnormality Androgenic activity

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A screening assay for thyroid hormone signaling disruption based on thyroid hormone-response gene expression analysis in the frog Pelophylax nigromaculatus

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Journal of Environmental Sciences 2015年第8期27卷 143-154页

作者： Yinfeng Zhang Yuanyuan Li zhanfen qin Huili Wang Jianzhong Li State Key Laboratory of Environmental Chemistry and Ecotoxicology Research Center for Eco-environmental Sciences Chinese Academy of Sciences Department of Environmental Bio-Technology Research Center for Eco-environmental Sciences Chinese Academy of Sciences

Amphibian metamorphosis provides a wonderful model to study the thyroid hormone（TH）signaling disrupting activity of environmental chemicals, with Xenopus laevis as the most commonly used species. This study aimed to establish a rapid and sensitive screening assay based on TH-response gene expression analysis using Pelophylax nigromaculatus, a native frog species distributed widely in East Asia, especially in China. To achieve this, five candidate TH-response genes that were sensitive to T3 induction were chosen as molecular markers, and T3 induction was determined as 0.2 nmol/L T3 exposure for 48 hr. The developed assay can detect the agonistic activity of T3 with a lowest observed effective concentration of 0.001 nmol/L and EC50 at around 0.118–1.229 nmol/L, exhibiting comparable or higher sensitivity than previously reported assays. We further validated the efficiency of the developed assay by detecting the TH signaling disrupting activity of tetrabromobisphenol A（TBBPA）, a known TH signaling disruptor. In accordance with previous reports, we found a weak TH agonistic activity for TBBPA in the absence of T3,whereas a TH antagonistic activity was found for TBBPA at higher concentrations in the presence of T3, showing that the P. nigromaculatus assay is effective for detecting TH signaling disrupting activity. Importantly, we observed non-monotonic dose-dependent disrupting activity of TBBPA in the presence of T3, which is difficult to detect with in vitro reporter gene assays. Overall, the developed P. nigromaculatus assay can be used to screen TH signaling disrupting activity of environmental chemicals with high sensitivity.

关键词： Thyroid hormone signaling disruptor Pelophylax nigromaculatus TH-response gene expression Screening assay Tetrabromobisphenol A

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Assessment of Bisphenol A(BPA) neurotoxicity in vitro with mouse embryonic stem cells

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Journal of Environmental Sciences 2015年第10期27卷 181-187页

作者： Nuoya Yin Xinglei Yao zhanfen qin Yuan-Liang Wang Francesco Faiola State Key Laboratory of Environmental Chemistry and Ecotoxicology Research Center for Eco-Environmental SciencesChinese Academy of Sciences Section of Molecular Biology University of California at San Diego

The adverse effects of environmental pollution on our well-being have been intensively studied with many in vitro and in vivo systems. In our group, we focus on stem cell toxicology due to the multitude of embryonic stem cell（ESC） properties which can be exerted in toxicity assays. In fact, ESCs can differentiate in culture to mimic embryonic development in vivo, or specifically to virtually any kind of somatic cells. Here, we used the toxicant Bisphenol A（BPA）, a chemical known as a hazard to infants and children, and showed that our stem cell toxicology system was able to efficiently recapitulate most of the toxic effects of BPA previously detected by in vitro system or animal tests. More precisely, we demonstrated that BPA affected the proper specification of germ layers during our in vitro mimicking of the embryonic development, as well as the establishment of neural ectoderm and neural progenitor cells.

关键词： Bisphenol A(BPA) Stem cell toxicology Neurotoxicity Embryonic stem cells(ESCs) Developmental toxicity Neural ectoderm

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