Brannen Kimberly C, Panzica-Kelly Julieta M, Danberry Tracy L, Augustine-Rauch Karen A
Discovery Toxicology, Bristol-Myers Squibb Company, Princeton, NJ, USA.
Birth Defects Res B Dev Reprod Toxicol. 2010 Feb;89(1):66-77. doi: 10.1002/bdrb.20223.
A zebrafish (Danio rerio) teratogenicity assay has been developed and evaluated for its ability to predict the teratogenic potential of chemicals.
Zebrafish embryos were dechorionated and then exposed to a test solution from 4-6 hours post-fertilization, and embryos or larvae were assessed up to 5 days post-fertilization (dpf) for viability and morphology. In preliminary experiments, the potential time points for assessment of compound-induced dysmorphology and general toxicity parameters were evaluated, and 5 dpf was found to be the optimum developmental stage for evaluation. Additionally, a morphological scoring system was devised to identify the developmental no-observed-adverse-effect level (NOAEL). For assay evaluation, 34 compounds with adequate in vivo developmental toxicity data were chosen. The compound set represented diversity in teratogenic potencies, structural classes, and pharmacologic targets. For 31 test compounds, each was evaluated over a concentration range, while 3 others were insufficiently aqueous-soluble to be fully tested. For each of the 31 tested compounds, the 5 dpf NOAEL was determined, and the concentration resulting in 25% lethality (LC25) was calculated by curve-fitting. Teratogenic potential of each compound was predicted based on the ratio of the LC25 to the NOAEL. LC25/NOAEL ratios of 10 or greater were considered predictive of teratogenicity.
The model successfully categorized 87% of the compounds as teratogens or non-teratogens, with only 2 false-positives (dimethyl phthalate and a Bristol-Myers Squibb (BMS) investigative compound) and 2 false-negatives (valproic acid and a BMS compound).
The results indicate that this assay is promising for screening compounds for teratogenic potential. Birth Defects Res (Part B) 89:66-77, 2010. (c) 2010 Wiley-Liss, Inc.
已开发出一种斑马鱼（Danio rerio）致畸性试验，并对其预测化学物质致畸潜力的能力进行了评估。
去除斑马鱼胚胎的卵膜，然后在受精后4至6小时将其暴露于测试溶液中，并在受精后5天（dpf）内评估胚胎或幼体的活力和形态。在初步实验中，评估了化合物诱导的畸形和一般毒性参数的潜在评估时间点，发现5 dpf是评估的最佳发育阶段。此外，设计了一种形态学评分系统来确定发育无观察到不良反应水平（NOAEL）。为了评估该试验，选择了34种具有充分体内发育毒性数据的化合物。该化合物集在致畸效力、结构类别和药理靶点方面具有多样性。对于31种测试化合物，每种化合物都在一个浓度范围内进行评估，而另外3种化合物水溶性不足，无法进行全面测试。对于31种测试化合物中的每一种，确定了5 dpf的NOAEL，并通过曲线拟合计算出导致25%致死率（LC25）的浓度。根据LC25与NOAEL的比值预测每种化合物的致畸潜力。LC25/NOAEL比值为10或更高被认为可预测致畸性。
该模型成功地将87%的化合物分类为致畸剂或非致畸剂，只有2例假阳性（邻苯二甲酸二甲酯和百时美施贵宝（BMS）的一种研究化合物）和2例假阴性（丙戊酸和一种BMS化合物）。
结果表明，该试验在筛选化合物致畸潜力方面具有前景。《出生缺陷研究（B部分）》89:66 - 77，2010年。（c）2010威利 - 利斯公司。
