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

Abstract

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.

ATP-binding cassette transporters at the zebrafish blood-brain barrier and the potential utility of the zebrafish as an in vivo model

“…In one comprehensive study, Kim et al [77] compared the zebrafish brain penetration of several compounds to penetration of the mouse brain. Among the compounds tested was loperamide, an analog of which has been explored for use as a PET imaging agent to measure P-gp function at the BBB [78] . The estimated partition coefficient for loperamide after oral administration in the zebrafish was found to increase by about 2-fold when coadministered with the P-gp inhibitor tariquidar, which was comparable to the 4-fold increase observed in mice.…”

“…Fischer et al [ 66 ] examined the accumulation of P-gp substrates rhodamine B and calcein-AM in whole embryo assays and demonstrated increased fluorescence in the embryos treated with the substrates and P-gp inhibitors compared to the embryos treated with the substrates alone. In a subsequent study, a number of compounds were also found to increase rhodamine B fluorescence in embryos at 72 hpf, potentially due to inhibition of zebrafish Abcb4 [ 79 ] . The cyanobacterial toxin microcystin-LR was also found to be a substrate of Abcb4 based on cytotoxicity assays with LLC-PK1 cells transfected with zebrafish abcb4 as well as microinjection of zebrafish embryos with abcb4 [ 80 ] .…”

Strategies to gain novel Alzheimer’s disease diagnostics and therapeutics using modulators of ABCA transporters

“…Several drugs and drug-like compounds have already been demonstrated to be pan-ABC transporter modulators interacting also with ABCA transporters. These drugs and drug-like compounds are, for example, cyclosporine A (9 targets of 4 subfamilies: ABCA1, 245 ABCB1, 20 ABCB4, 552 ABCB11, 553 ABCC1–2, 24 , 554 ABCC10, 26 and ABCG1–2 555 , 556 ), glibenclamide (8 targets of 4 subfamilies: ABCA1, 270 ABCB11, 553 ABCC1, 24 ABCC5, 557 ABCC7–9, 558 - 560 and ABCG2 554 ), imatinib (6 targets of 4 subfamilies: ABCA3, 426 ABCB1, 561 ABCB11, 553 ABCC1, 561 ABCC10, 561 and ABCG2 561 ), probenecid (8 targets of 2 subfamilies: ABCA8, 222 ABCC1–6, 24 , 26 , 562 - 564 ABCC10 565 ), verapamil (9 targets of 4 subfamilies: ABCA8, 222 ABCB1, 20 ABCB4–5, 552 , 566 ABCB11, 567 ABCC1, 24 ABCC4, 568 ABCC10, 565 and ABCG2 554 ), and verlukast (11 targets of 4 subfamilies: ABCA8, 222 ABCB4, 552 ABCB11, 553 ABCC1–5, 24 , 554 , 557 , 564 , 569 ABCC10–11, 26 , 570 ABCG2 554 ). In silico analyses with verapamil and verlukast supported the notion of addressing the multitarget binding site in ABCA7.…”

Scaffold fragmentation and substructure hopping reveal potential, robustness, and limits of computer-aided pattern analysis (C@PA)

“…Multitargeting is a promising approach to explore under-studied ABC transporters by targeting similar or mutually overlapping binding sites [15] , [73] , [74] . Several pharmacological drugs have already been revealed as (weak) pan-ABC transporter inhibitors (=inhibiting several ABC transporters simultaneously), as for example, benzbromarone ( 1 ; ABCB1 [75] , ABCB11 [20] , ABCC1-6 [23] , [24] , [58] , [76] , [77] , and ABCG2 [75] ), cyclosporine A ( 2 ; ABCA1 [27] , ABCB1 [16] , ABCB4 [78] , ABCB11 [20] , ABCC1–2 [23] , [58] , ABCC10 [24] , ABCG1–2 [75] , [79] ), glibenclamide (glyburide, 3 ; ABCA1 [29] , ABCB11 [20] , ABCC1 [23] , ABCC5 [80] , ABCC7–9 [81] , [82] , [83] , ABCG2 [58] ), probenecid ( 4 ; ABCA8 [84] , ABCC1–6 [23] , [24] , [85] , [86] , [87] , ABCC10 [88] ), verapamil ( 5 ; ABCA8 [84] , ABCB1 [16] , ABCB4–5 [54] , [78] , ABCB11 [89] , ABCC1 [23] , ABCC4 [90] , ABCC10 [88] , ABCG2 [58] ), or verlukast (MK571, 6 ; ABCA8 [84] , ABCB4 [78] , ABCB11 [20] , ABCC1–5 [23] , [58] , [80] , [87] , [91] , ABCC10–11 [24] , [92] , ABCG2 [58] ). Fig.…”