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Molecular Pharmacology

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Research ArticleArticle

Identification of Zebrafish ARNT1 Homologs: 2,3,7,8-Tetrachlorodibenzo-p-dioxin Toxicity in the Developing Zebrafish Requires ARNT1

Amy L. Prasch, Robert L. Tanguay, Vatsal Mehta, Warren Heideman and Richard E. Peterson
Molecular Pharmacology March 2006, 69 (3) 776-787; DOI: https://doi.org/10.1124/mol.105.016873
Amy L. Prasch
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Robert L. Tanguay
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Vatsal Mehta
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Warren Heideman
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Richard E. Peterson
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Abstract

To use the zebrafish (Danio rerio) as a model to study 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) developmental toxicity, it is essential to know which proteins are involved in mediating toxicity. Previous work has identified zfAHR2 as the receptor that binds TCDD mediating downstream responses. Although zfARNT2b can form a functional heterodimer with zfAHR2 in vitro, zfarnt2 null mutants show no protection against endpoints of TCDD developmental toxicity, demonstrating that zfARNT2b cannot be the physiological dimerization partner for zfAHR2 mediating responses to TCDD in zebrafish embryos. The purpose of the current study was to identify an alternate dimerization partner(s) for zfAHR2 that may function to mediate TCDD developmental toxicity. By searching zebrafish genomic sequence and using the polymerase chain reaction-based rapid amplification of cDNA ends technique, three forms of cDNA that seem to be alternate mRNA splice variants of a zebrafish homolog of ARNT1 were detected. Analysis of the zfARNT1 proteins in vitro demonstrates that the two longest forms of zfARNT1, zfARNT1b and zfARNT1c, can form functional heterodimers with zfAHR2. However, the shortest form, zfARNT1a, seems to be nonfunctional with zfAHR2 in vitro. To determine whether a zfARNT1 protein functions with zfAHR2 in vivo, a morpholino targeted against the 5′ end of zfARNT1 (zfarnt1-MO) was used. Injection of the zfarnt1-MO before TCDD treatment significantly decreases the induction of zfCYP1A mRNA and protein. In addition, zfarnt1 morphants show complete protection against TCDD-induced pericardial edema and show partial protection against reduced blood flow and craniofacial malformations caused by TCDD, demonstrating the role of zfARNT1 proteins in mediating these responses.

  • Received July 19, 2005.
  • Accepted November 9, 2005.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 69 (3)
Molecular Pharmacology
Vol. 69, Issue 3
1 Mar 2006
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Research ArticleArticle

Identification of Zebrafish ARNT1 Homologs: 2,3,7,8-Tetrachlorodibenzo-p-dioxin Toxicity in the Developing Zebrafish Requires ARNT1

Amy L. Prasch, Robert L. Tanguay, Vatsal Mehta, Warren Heideman and Richard E. Peterson
Molecular Pharmacology March 1, 2006, 69 (3) 776-787; DOI: https://doi.org/10.1124/mol.105.016873

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Research ArticleArticle

Identification of Zebrafish ARNT1 Homologs: 2,3,7,8-Tetrachlorodibenzo-p-dioxin Toxicity in the Developing Zebrafish Requires ARNT1

Amy L. Prasch, Robert L. Tanguay, Vatsal Mehta, Warren Heideman and Richard E. Peterson
Molecular Pharmacology March 1, 2006, 69 (3) 776-787; DOI: https://doi.org/10.1124/mol.105.016873
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