Journal
FEBS LETTERS
Volume 595, Issue 6, Pages 828-843Publisher
WILEY
DOI: 10.1002/1873-3468.14015
Keywords
ABC efflux transporter; ATPase activity; chemosensitizer; dye accumulation assay; zebrafish Abcb4; zebrafish embryo
Funding
- German Academic Exchange Service (Deutscher Akademischer Austauschdienst-DAAD, ALEARG program)
- German Federal Environmental Foundation (Deutsche Bundesstiftung Umwelt-DBU)
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An automated microscopy-based rhodamine B dye accumulation assay was established to evaluate the activity of multixenobiotic efflux transporter in zebrafish embryos, confirming the function of Abcb4 as an efflux transporter in zebrafish and showing potential for screening environmental pollutants and chemicals.
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.
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