Chemical effects on dye efflux activity in live zebrafish embryos and on zebrafish Abcb4 ATPase activity,FEBS Letters

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Chemical effects on dye efflux activity in live zebrafish embryos and on zebrafish Abcb4 ATPase activity
FEBS Letters ( IF 3.0 ) Pub Date : 2020-12-21 , DOI: 10.1002/1873-3468.14015
Flavia Bieczynski _{1,

2} , Kathleen Burkhardt-Medicke ₂ , Carlos M Luquet ₃ , Stefan Scholz ₂ , Till Luckenbach ₂

Affiliation

ATP binding cassette (ABC) transporter proteins include efflux pumps that confer multixenobiotic resistance to zebrafish embryos, a valuable toxico/pharmacological model. Here, we established an automated microscopy-based rhodamine B dye accumulation assay in which enhanced dye accumulation in live zebrafish embryos indicates inhibition of multixenobiotic efflux transporter activity. Twenty structurally divergent known substrates and/or inhibitors of human ABC transporters and environmentally relevant compounds were examined using this assay and the ATPase activity of recombinant zebrafish Abcb4 as readouts. These two assays confirmed that Abcb4 functions as an efflux transporter in zebrafish, whereas they gave discordant results for some of the tested substances. The dye accumulation assay in zebrafish embryos could be useful to screen environmental pollutants and other chemicals for efflux transporter interaction in a medium-throughput fashion.

中文翻译：

对活斑马鱼胚胎染料外排活性和斑马鱼 Abcb4 ATPase 活性的化学影响

ATP 结合盒 (ABC) 转运蛋白包括外排泵，可赋予斑马鱼胚胎多异生物抗性，这是一种有价值的毒理学/药理学模型。在这里，我们建立了一种基于自动显微镜的罗丹明 B 染料积累测定，其中活斑马鱼胚胎中增强的染料积累表明对多异生物外排转运蛋白活性的抑制。使用该测定法和重组斑马鱼 Abcb4 的 ATP 酶活性作为读数检查了 20 种结构不同的已知底物和/或人类 ABC 转运蛋白和环境相关化合物的抑制剂。这两项测定证实了 Abcb4 在斑马鱼中作为外排转运蛋白发挥作用，而对于一些测试物质，它们给出了不一致的结果。

更新日期：2020-12-21

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