胚胎期双酚A暴露对斑马鱼发育及神经行为的影响
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摘要
目的·运用斑马鱼模型初步探讨胚胎期双酚A(bisphenol A,BPA)暴露对斑马鱼发育和神经行为的影响。方法·对受精后小时数(hours post-fertilization,hpf)为4的正常斑马鱼受精卵以不同浓度BPA(0、2.5、25、250、2 500μg/L)作染毒处理;观察并记录斑马鱼存活率、畸形率和孵化率,以及斑马鱼胚胎24 hpf自主运动和48 hpf心率,并对发育至144 hpf的斑马鱼幼鱼进行神经行为学检测。结果·各浓度BPA均未影响斑马鱼的存活率、畸形率和孵化率。暴露于2 500μg/L浓度BPA的斑马鱼胚胎24 hpf自主运动和48 hpf心率低于对照组(P=0.000),发育至144 hpf的幼鱼运动总距离和平均速度降低(P=0.000)。暴露于250μg/L和2 500μg/L浓度BPA的斑马鱼幼鱼在强光刺激惊恐反射检测中运动行为改变,主要表现为在强光刺激下运动总距离减少(P<0.05)。结论·胚胎期BPA暴露对斑马鱼具有一定的发育毒性和神经毒性。
Objective · To investigate the effects of bisphenol A(BPA) on the development and neurobehavior of zebrafish. Methods · The zebrafish fertilized eggs that developed to 4 hpf(hours post-fertilization) were treated with different concentrations of BPA(0, 2.5, 25, 250, and 2 500 μg/L).Survival rates, malformation rates, hatching rates, 24 hpf autonomic movement, and 48 hpf heart rates in the groups were observed and recorded. And the neurobehavioral test of larval zebrafish that developed to 144 hpf was performed. Results · The BPA exposure did not affect the survival rates,malformation rates and hatching rates of zebrafish. Compared with the control group, the autonomic movement in 24 hpf and heart rate in 48 hpf of zebrafish embryos that exposed to 2 500 μg/L BPA decreased(P=0.000), and the total distance and average speed of larval zebrafish that developed to 144 hpf also decreased(P=0.000). The behavior of larval zebrafish exposed to 250 μg/L and 2 500 μg/L BPA changed on light-evoked startle escape response activity, mainly including the decrease of total distance under strong light stimulation(P<0.05). Conclusion · BPA exposure during embryonic phase has developmental toxicity and neurotoxicity on zebrafish.
Objective · To investigate the effects of bisphenol A(BPA) on the development and neurobehavior of zebrafish. Methods · The zebrafish fertilized eggs that developed to 4 hpf(hours post-fertilization) were treated with different concentrations of BPA(0, 2.5, 25, 250, and 2 500 μg/L).Survival rates, malformation rates, hatching rates, 24 hpf autonomic movement, and 48 hpf heart rates in the groups were observed and recorded. And the neurobehavioral test of larval zebrafish that developed to 144 hpf was performed. Results · The BPA exposure did not affect the survival rates,malformation rates and hatching rates of zebrafish. Compared with the control group, the autonomic movement in 24 hpf and heart rate in 48 hpf of zebrafish embryos that exposed to 2 500 μg/L BPA decreased(P=0.000), and the total distance and average speed of larval zebrafish that developed to 144 hpf also decreased(P=0.000). The behavior of larval zebrafish exposed to 250 μg/L and 2 500 μg/L BPA changed on light-evoked startle escape response activity, mainly including the decrease of total distance under strong light stimulation(P<0.05). Conclusion · BPA exposure during embryonic phase has developmental toxicity and neurotoxicity on zebrafish.
引文
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[13]Zhao F,Wei P,Wang J,et al.Estrogenic effects associated with bisphenol A exposure in male zebrafish(Danio rerio)is associated with changes of endogenous 17β-estradiol and gene specific DNA methylation levels[J].Gen Comp Endocrinol,2017,252:27-35.
[14]Chan WK,Chan KM.Disruption of the hypothalamic-pituitary-thyroid axis in zebrafish embryo-larvae following waterborne exposure to BDE-47,TBBPAand BPA[J].Aquat Toxicol,2012,108:106-111.
[15]Fowden AL,Forhead AJ.Hormones as epigenetic signals in developmental programming[J].Exp Physiol,2009,94(6):607-625.
[2]李敏.中国双酚A市场分析及前景展望[J].中国石油和化工经济分析,2017(11):55-57.
[3]Mustieles V,Pérez-Lobato R,Olea N,et al.Bisphenol A:human exposure and neurobehavior[J].Neurotoxicology,2015,49:174-184.
[4]Silveira PP,Portella AK,Goldani MZ,et al.Developmental origins of health and disease(DOHaD)[J].J Pediatr(Rio J),2007,83(6):494-504.
[5]Bambino K,Chu J.Zebrafish in toxicology and environmental health[J].Curr Top Dev Biol,2017,124:331-367.
[6]邹苏琪,殷梧,杨昱鹏,等.斑马鱼行为学实验在神经科学中的应用[J].生物化学与生物物理进展,2009,36(1):5-12.
[7]Pan R,Wang CF,Shi R,et al.Prenatal bisphenol A exposure and early childhood neurodevelopment in Shandong,China[J].Int J Hyg Environ Health,2019.DOI:10.1016/j.ijheh.2019.03.002.
[8]Mu X,Huang Y,Li X,et al.Developmental effects and estrogenicity of bisphenol A alternatives in a zebrafish embryo model[J].Environ Sci Technol,2018,52(5):3222-3231.
[9]郭依晨,韩静静,严锐,等.双酚A胚胎期暴露对斑马鱼的发育毒性及运动行为的影响[J].毒理学杂志,2018,32(2):71-74.
[10]杨丽华,史奇朋,周炳升.双酚A对斑马鱼幼鱼神经递质和神经行为的影响[J].生态毒理学报,2017,12(3):162-169.
[11]Fraser TWK,Khezri A,Lewandowska-Sabat AM,et al.Endocrine disruptors affect larval zebrafish behavior:testing potential mechanisms and comparisons of behavioral sensitivity to alternative biomarkers[J].Aquat Toxicol,2017,193:128-135.
[12]Fraser TWK,Khezri A,Jusdado JGH,et al.Toxicant induced behavioural aberrations in larval zebrafish are dependent on minor methodological alterations[J].Toxicol Lett,2017,276:62-68.
[13]Zhao F,Wei P,Wang J,et al.Estrogenic effects associated with bisphenol A exposure in male zebrafish(Danio rerio)is associated with changes of endogenous 17β-estradiol and gene specific DNA methylation levels[J].Gen Comp Endocrinol,2017,252:27-35.
[14]Chan WK,Chan KM.Disruption of the hypothalamic-pituitary-thyroid axis in zebrafish embryo-larvae following waterborne exposure to BDE-47,TBBPAand BPA[J].Aquat Toxicol,2012,108:106-111.
[15]Fowden AL,Forhead AJ.Hormones as epigenetic signals in developmental programming[J].Exp Physiol,2009,94(6):607-625.
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