Abstract：
Detecting the toxic effects of chemicals on reproduction and development without using mammalian animal models is crucial in the exploitation of pharmaceuticals for human use. Zebrafish are a promising animal model for investigating pharmacological effects and toxicity during vertebrate development. Several studies have suggested the use of zebrafish embryos for the assessment of malformations or embryo-fetal lethality (MEFL). However, a reproducible protocol as a standard for the zebrafish MEFL test method that fulfills global requests has not been established based on the International Council of Harmonisation (ICH) S5 (R3) guidelines. To establish such a toxicity test method, we developed a new and easy protocol to detect MEFL caused by chemicals, especially those with teratogenic potential, using fertilized zebrafish eggs (embryos) within 5 days of development. Our toxicity test trials using the same protocol in two to four different laboratories corroborated the high inter-laboratory reproducibility. Our test method enabled the detection of 18 out of 22 test compounds that induced rat MEFL. Thus, the prediction rate of our zebrafish test method for MEFL was almost 82% compared with that of rat MEFL. Collectively, our study proposes the establishment of an easy and reproducible protocol for the zebrafish MEFL test method for reproductive and developmental toxicity that meets ICH guideline S5 (R3), which can be further considered in combination with information from other sources for regulatory use.
摘要：
在不使用哺乳动物动物模型的情况下检测化学物质对生殖和发育的毒性作用对于开发人类使用的药物至关重要。斑马鱼是研究脊椎动物发育过程中的药理作用和毒性的有前途的动物模型。一些研究建议使用斑马鱼胚胎来评估畸形或胚胎-胎儿致死率（MEFL）。然而,根据国际协调理事会(ICH)S5(R3)指南,尚未建立可重复的方案作为满足全球要求的斑马鱼MEFL测试方法的标准.为了建立这样的毒性测试方法，我们开发了一种新的简单的协议来检测由化学物质引起的MEFL，尤其是那些具有致畸潜力的人，在发育的5天内使用受精的斑马鱼卵（胚胎）。我们在两到四个不同实验室中使用相同方案进行的毒性测试试验证实了高度的实验室间可重复性。我们的测试方法能够检测到22种诱导大鼠MEFL的测试化合物中的18种。因此，我们的斑马鱼测试方法对MEFL的预测率与大鼠MEFL相比几乎为82％。总的来说，我们的研究提出了建立一个容易和可重复的方案，用于斑马鱼MEFL测试方法的生殖和发育毒性，符合ICH指南S5（R3），可以结合其他来源的信息进一步考虑，以供监管使用。
