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Advancing the Zebrafish Embryo Test for Endocrine Disruptor Screening Using Micro-Injection: Ethinyl Estradiol as a Case Study

Advancing the Zebrafish Embryo Test for Endocrine Disruptor Screening Using Micro-Injection: Ethinyl Estradiol as a Case Study

Ellen D G Michiels, Lucia Vergauwen, Foon Yin Lai, Raewyn M Town, Adrian Covaci, Alexander L N van Nuijs, Steven J Van Cruchten, Dries Knapen

Environ Toxicol Chem. 2019 Mar;38(3):533-547.

PMID: 30569562

Abstract:

Fish (embryo) toxicity test guidelines are mostly based on aquatic exposures. However, in some cases, other exposure routes can be more practical and relevant. Micro-injection into the yolk of fish embryos could offer a particular advantage for administering hydrophobic compounds, such as many endocrine disruptors. Single-dose micro-injection was compared with continuous aquatic exposure in terms of compound accumulation and biological responses. 17α-Ethinyl estradiol (EE2) was used as a model compound. First, the optimal solvent and droplet size were optimized, and needle variation was assessed. Next, biological endpoints were evaluated. The accumulated internal dose of EE2 decreased over time in both exposure scenarios. Estrogen receptor activation was concentration/injected dose dependent, increased daily, and was related to esr2b transcription. Transcription of vitellogenin 1 (vtg1) and brain aromatase (cyp19a1b) was induced in both scenarios, but the cyp19a1b transcription pattern differed between routes. Injection caused an increase in cyp19a1b transcripts from 48 hours post fertilization (hpf) onward, whereas after aquatic exposure the main increase occurred between 96 and 120 hpf. Some malformations only occurred after injection, whereas others were present for both scenarios. We conclude that responses can differ between exposure routes and therefore micro-injection is not a direct substitute for, but can be complementary to aquatic exposure. Nevertheless, vtg1and cyp19a1b transcription and estrogen receptor activation are suitable biomarkers for endocrine disruptor screening in both scenarios. Environ Toxicol Chem 2019;38:533-547. © 2018 SETAC.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP38002185	Ethinyl estradiol Related Compound A		38002-18-5	Price

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