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Zebrafish Oatp-mediated transport of microcystin congeners

  • Molecular Toxicology
  • Published:
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Abstract

Microcystins (MC), representing >100 congeners being produced by cyanobacteria, are a hazard for aquatic species. As MC congeners vary in their toxicity, the congener composition of a bloom primarily dictates the severity of adverse effects and appears primarily to be governed by toxicokinetics, i.e., whether transport of MCs occurs via organic anion-transporting polypeptides (Oatps). Differences in observed MC toxicity in various fish species suggest differential expression of Oatp subtypes leading to varying tissue distribution of the very same MC congener within different species. The objectives of this study were the functional characterization and analysis of the tissue distribution of Oatp subtypes in zebrafish (Danio rerio) as a surrogate model for cyprinid fish. Zebrafish Oatps (zfOatps) were cloned, and the organ distribution was determined at the mRNA level. zfOatps were transiently expressed in HEK293 cells for functional characterization using the Oatp substrates estrone-3-sulfate, taurocholate and methotrexate and specific MC congeners (MC-LR, MC-RR, MC-LF and MC-LW). Novel zfOatp isoforms were isolated. Among these isoforms, the organ-specific expression of zfOatp1d1 and of members of the zfOatp1f subfamily was identified. At the functional level, zfOatp1d1, zfOatp1f2, zfOatp1f3 and zfOatp1f4 transported at least one of the Oatp substrates, and zfOatp1d1, zfOatp1f2 and zfOatp1f4 were shown to transport MC congeners. MC-LF and MC-LW were generally transported faster than MC-LR and MC-RR. The subtype-specific expression of zfOatp1d1 and of members of the zfOatp1f subfamily as well as differences in the transport of MC congeners could explain the MC congener-dependent differences in toxicity in cyprinids.

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Abbreviations

OATP:

Organic anion-transporting polypeptide (in humans and other mammalians)

MC:

Microcystin

MC-LR:

Microcystin-LR

MC-RR:

Microcystin-RR

MC-LW:

Microcystin-LW

MC-LF:

Microcystin-LF

ser/thrPP:

Ser/Thr protein phosphatases

PP1:

Protein phosphatase 1

Oatp:

Organic anion-transporting polypeptide (in fish)

zf:

Zebrafish

h:

Human

m:

Mouse

ORF:

Open reading frame

TCA:

Taurocholic acid

E3S:

Estrone sulfate ammonium salt

MTX:

Methotrexate disodium salt

TMD:

Transmembrane domain

HEK:

Human embryonic kidney

HeLa:

Henrietta Lacks

GFP:

Green fluorescent protein

slco:

Solute carrier organic anion transporter family

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Acknowledgments

The financial support from the DFG Center Grant, International Max Planck Research School for Organismal Biology (IMPRS), Marie Curie International Research Staff Exchange Scheme Fellowship (PIRSES-GA-2011-295223), the National Center for Research Resources (RR021940) and the National Institute of General Medical Sciences (GM077336 and GM103549) of the National Institutes of Health are acknowledged.

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Authors and Affiliations

  1. Human and Environmental Toxicology, Department of Biology, University of Konstanz, PO BOX 662, 78457, Constance, Germany

    Konstanze Steiner, Lisa Zimmermann & Daniel Dietrich

  2. Pharmacology, Toxicology and Therapeutics, The University of Kansas Medical Center, Kansas City, KS, USA

    Bruno Hagenbuch

Authors
  1. Konstanze Steiner
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  2. Lisa Zimmermann
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  3. Bruno Hagenbuch
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  4. Daniel Dietrich
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Correspondence to Daniel Dietrich.

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Steiner, K., Zimmermann, L., Hagenbuch, B. et al. Zebrafish Oatp-mediated transport of microcystin congeners. Arch Toxicol 90, 1129–1139 (2016). https://doi.org/10.1007/s00204-015-1544-3

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  • DOI: https://doi.org/10.1007/s00204-015-1544-3

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