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

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JARQUE ORTIZ Sergio FETTER Eva VENEMAN Wouter J. SPAINK Herman P. PERAVALI Ravindra STRAEHLE Uwe SCHOLZ Stefan

Rok publikování 2018
Druh Článek v odborném periodiku
Časopis / Zdroj Plos one
Fakulta / Pracoviště MU

Přírodovědecká fakulta

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Doi http://dx.doi.org/10.1371/journal.pone.0203087
Klíčová slova THYROID-GLAND; TOXICITY TEST; FISH; EXPRESSION; DISRUPTION; POLLUTANTS; CHEMICALS; EXPOSURE; MODEL; ASSAY
Popis 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.
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