An automated screening method for detecting compounds with goitrogenic activity using transgenic zebrafish embryos

Sergio Jarque; Eva Fetter; Wouter J Veneman; Herman P Spaink; Ravindra Peravali; Uwe Strähle; Stefan Scholz

doi:10.1371/journal.pone.0203087

An automated screening method for detecting compounds with goitrogenic activity using transgenic zebrafish embryos

PLoS One. 2018 Aug 29;13(8):e0203087. doi: 10.1371/journal.pone.0203087. eCollection 2018.

Authors

Sergio Jarque¹, Eva Fetter², Wouter J Veneman³, Herman P Spaink³, Ravindra Peravali⁴, Uwe Strähle⁴, Stefan Scholz²

Affiliations

¹ RECETOX, Faculty of Science, Masaryk University, Brno, Czech Republic.
² Department of Bioanalytical Ecotoxicology, Helmholtz Centre for Environmental Research-UFZ, Leipzig, Germany.
³ Department of Animal Sciences and Health, Institute of Biology, Leiden University, Leiden, The Netherlands.
⁴ Institute of Toxicology and Genetics, Karlsruhe Institute of Technology, Karlsruhe, Germany.

Abstract

The knowledge on environmentally relevant chemicals that may interfere with thyroid signaling is scarce. Here, we present a method for the screening of goitrogens, compounds that disrupt the thyroid gland function, based on the automatic orientation of zebrafish in a glass capillary and a subsequent imaging of reporter gene fluorescence in the thyroid gland of embryos of the transgenic zebrafish line tg(tg:mCherry). The tg(tg:mCherry) reporter gene indicates a compensatory upregulation of thyroglobulin, the thyroid hormone precursor, in response to inhibition of thyroid hormone synthesis. Fish embryos were exposed to a negative control compound (3,4-dichloroaniline), or a concentration series of known goitrogenic compounds (resorcinol, methimazole, potassium perchlorate, 6-propyl-2-thiouracil, ethylenethiourea, phloroglucinol, pyrazole) with maximum exposure concentration selected based on mortality and/or solubility. Exposure to 3,4-dichloroaniline decreased the fluorescence signal. All goitrogenic compounds exhibited clear concentration-dependent inductions of reporter fluorescence 1.4 to 2.6 fold above control levels. Concentration-response modelling was used to calculate goitrogenic potencies based on EC50 values. The new automated method offers an efficient screening approach for goitrogenic activity.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Animals, Genetically Modified
Antithyroid Agents / pharmacology*
Automation, Laboratory*
Dose-Response Relationship, Drug
Drug Evaluation, Preclinical / methods*
Embryo, Nonmammalian / drug effects
Embryo, Nonmammalian / metabolism
Hydrophobic and Hydrophilic Interactions
Image Processing, Computer-Assisted
Luminescent Proteins / genetics
Luminescent Proteins / metabolism
Microscopy, Fluorescence
Thyroid Gland / drug effects
Thyroid Gland / metabolism
Zebrafish

Substances

Antithyroid Agents
Luminescent Proteins

Grants and funding

The work of Eva Fetter was supported by a scholarship of the German Environmental Scholarship foundation. Sergio Jarque was supported by the Czech Ministry of Education (LO1214) and the Program of “Employment of Newly Graduated Doctors of Science for Scientific Excellence” (Grant number CZ.1.07/2.3.00/30.0009) co-financed from European Social Fund and the State Budget of the Czech Republic. All other authors did not receive specific fundings for their contribution. We gratefully acknowledge access to the platform CITEPro (Chemicals in the Terrestrial Environment Profiler) funded by the Helmholtz Association. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.