Zebrafish: A preclinical model for drug screening
Zebrafish: A preclinical model for drug screening
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DOI：
10.1089/154065802761001293
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发表时间：
2002-11-01
影响因子：
1.8
通讯作者：
McGrath, P
中科院分区：
文献类型：
--
作者：
Parng, C;Seng, WL;McGrath, P
斑马鱼胚胎已成为评估药物作用的重要脊椎动物模型。它非常适合遗传学，胚胎学，发育和细胞生物学的研究。斑马鱼胚胎具有独特的特征，包括易于维护和药物管理，生殖周期短，以及允许对发育中的细胞和器官进行视觉评估的透明度。由于这些优点，斑马鱼生物测定比小鼠测定更便宜和更快，并且适合于大规模药物筛选。在这里，我们描述了使用斑马鱼生物测定评估毒性，血管生成和细胞凋亡。使用18种化学物质，我们证明了斑马鱼的毒性反应，致畸作用和LC 50与小鼠的结果相当。在没有复杂处理的透明动物中观察到化合物对各种器官（包括心脏、脑、肠、胰腺、软骨、肝和肾）的作用，证明了使用斑马鱼胚胎的毒性测定的效率。使用内源性碱性磷酸酶染色和全动物酶测定，我们证明了SU5416和flavopiridol，化合物显示在哺乳动物中具有抗血管生成作用，抑制斑马鱼血管生长，这种生物测定法适用于使用96孔酶标仪进行高通量筛选。我们还证明，在体内吖啶橙子染色可用于可视化与布雷菲德菌素A，新霉素，或半胱天冬酶抑制剂处理的胚胎中的凋亡事件。在体内染色后，吖啶橙子可被提取并使用荧光酶标仪定量，从而提供用于调节细胞凋亡的试剂的筛选系统。
The zebrafish embryo has become an important vertebrate model for assessing drug effects. It is well suited for studies in genetics, embryology, development, and cell biology. Zebrafish embryos exhibit unique characteristics, including ease of maintenance and drug administration, short reproductive cycle, and transparency that permits visual assessment of developing cells and organs. Because of these advantages, zebrafish bioassays are cheaper and faster than mouse assays, and are suitable for large-scale drug screening. Here we describe the use of zebrafish bioassays for assessing toxicity, angiogenesis, and apoptosis. Using 18 chemicals, we demonstrated that toxic response, teratogenic effects, and LC50 in zebrafish are comparable to results in mice. The effects of compounds on various organs, including the heart, brain, intestine, pancreas, cartilage, liver, and kidney, were observed in the transparent animals without complicated processing, demonstrating the efficiency of toxicity assays using zebrafish embryos. Using endogenous alkaline phosphatase staining and a whole-animal enzyme assay, we demonstrated that SU5416 and flavopiridol, compounds shown to have antiangiogenic effects in mammals, inhibit blood vessel growth in zebrafish, and this bioassay is suitable for high-throughput screening using a 96-well microplate reader. We also demonstrated that in vivo acridine orange staining can be used to visualize apoptotic events in embryos treated with brefeldin A, neomycin, or caspase inhibitors. After in vivo staining, acridine orange can be extracted and quantitated using a fluorescence microplate reader, providing a screening system for agents that modulate apoptosis.