转基因斑马鱼模型已成功用于环境污染物的生物监测和风险评估研究。包括异雌激素,杀虫剂,和重金属。我们采用斑马鱼幼虫(转基因SR4G系)和皮质醇诱导的绿色荧光蛋白报告基因(eGFP)作为模型来检测暴露于具有环境影响的化合物后的应激反应,包括双酚A(BPA),vinclozolin(VIN),和氟西汀(FLX)。皮质醇,荧光信号,在斑马鱼幼虫的均质池中测量eGFP和11个靶向基因的mRNA水平,每个终点有六个实验重复。根据它们与应激轴和基因的立即早期反应类别的关联选择11个靶基因。用氢化可的松(CORT)和地塞米松(DEX)作为阳性和阴性对照,分别。所有测量均在两个无应力和应力条件下进行,使用标准化的净处理作为应力源。在皮质醇水平和eGFPmRNA水平之间观察到显著的正线性相关(r>0.9)。基于未应激和应激幼虫中的eGFPmRNA水平,训练两个预测模型(随机森林和逻辑回归)。这两种模型都可以正确预测暴露于BPA后的钝化应激反应,VIN,FLX和阴性对照,DEX.这些模型的阴性预测值(NPV)为100%。当基于11个评估基因的mRNA水平训练预测模型时,观察到类似的NPV。全身荧光强度信号的测量对于检测钝化的应激反应并不显著。我们的发现支持使用SR4G转基因幼虫作为体内生物监测模型来筛选化学物质的应激破坏潜力。这很重要,因为越来越多的证据表明,短暂暴露于环境污染物会改变几代人的应激反应和关键应对行为。
Transgenic zebrafish models have been successfully used in biomonitoring and risk assessment studies of environmental pollutants, including xenoestrogens, pesticides, and heavy metals. We employed zebrafish larva (transgenic SR4G line) with a cortisol-inducible green fluorescence protein reporter (eGFP) as a model to detect stress responses upon exposure to compounds with environmental impact, including bisphenol A (BPA), vinclozolin (VIN), and fluoxetine (FLX). Cortisol, fluorescence signal, and mRNA levels of eGFP and 11 targeted genes were measured in a homogenized pool of zebrafish larvae, with six experimental replicates for each endpoint. Eleven targeted genes were selected according to their association with stress-axis and immediate early response class of genes. Hydrocortisone (CORT)and dexamethasone (DEX) were used as positive and negative controls, respectively. All measurements were done in two unstressed and stressed condition using standardized net handling as the stressor. A significant positive linear correlation between cortisol levels and eGFP mRNA levels was observed (r> 0.9). Based on eGFP mRNA levels in unstressed and stressed larvae two predictive models were trained (Random Forest and Logistic Regression). Both these models could correctly predict the blunted stress response upon exposure to BPA, VIN, FLX and the negative control, DEX. The negative predictive value (NPV) of these models were 100%. Similar NPV was observed when the predictive models trained based on the mRNA levels of the eleven assessed genes. Measurement of whole-body fluorescence intensity signal was not significant to detect blunted stress response. Our findings support the use of SR4G transgenic larvae as an in vivo biomonitoring model to screen chemicals for their stress-disrupting potentials. This is important because there is increasing evidence that brief exposures to environmental pollutants modify the stress response and critical coping behaviors for several generations.