A Multiwell-Based Assay for Screening Thyroid Hormone Signaling Disruptors Using thibz Expression as a Sensitive Endpoint in Xenopus laevis

There is a need for rapidly screening thyroid hormone (TH) signaling disruptors in vivo considering the essential role of TH signaling in vertebrates. We aimed to establish a rapid in vivo screening assay using based on the T3-induced metamorphosis assay we established previously, as well as the Ele...

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Bibliographic Details
Published in:Molecules (Basel, Switzerland) Switzerland), 2022-01, Vol.27 (3), p.798
Main Authors: Li, Jinbo, Li, Yuanyuan, Zhu, Min, Song, Shilin, Qin, Zhanfen
Format: Article
Language:English
Subjects:
Alanine - analogs & derivatives
Alanine - pharmacology
Animals
Assaying
Bisphenol A
Endocrine Disruptors - pharmacology
Gene expression
Gene Expression - drug effects
Halogenated Diphenyl Ethers - pharmacology
Helper cells
High-Throughput Screening Assays - methods
Kinases
Lymphocytes T
Metamorphosis
multiwell plate
screening assay
Signal Transduction - drug effects
Tetrabromobisphenol A
thibz gene
Thyroid
Thyroid gland
thyroid hormone
Thyroid Hormones - metabolism
Triiodothyronine
Triiodothyronine - pharmacology
Vertebrates
Xenopus
Xenopus laevis
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Summary:There is a need for rapidly screening thyroid hormone (TH) signaling disruptors in vivo considering the essential role of TH signaling in vertebrates. We aimed to establish a rapid in vivo screening assay using based on the T3-induced metamorphosis assay we established previously, as well as the Eleutheroembryonic Thyroid Assay (XETA). Stage 48 tadpoles were treated with a series of concentrations of T3 in 6-well plates for 24 h and the expression of six TH-response genes was analyzed for choosing a proper T3 concentration. Next, bisphenol A (BPA) and tetrabromobisphenol A (TBBPA), two known TH signaling disruptors, were tested for determining the most sensitive TH-response gene, followed by the detection of several suspected TH signaling disruptors. We determined 1 nM as the induction concentration of T3 and expression as the sensitive endpoint for detecting TH signaling disruptors given its highest response to T3, BPA, and TBBPA. And we identified betamipron as a TH signaling agonist, and 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) as a TH signaling antagonist. Overall, we developed a multiwell-based assay for rapidly screening TH signaling disruptors using expression as a sensitive endpoint in .
ISSN:1420-3049
1420-3049
DOI:10.3390/molecules27030798