In Vitro and in Vivo Antiestrogenic Effects of Polycyclic Musks in Zebrafish

• Published in: Environmental science & technology Vol. 38; no. 4; pp. 997 - 1002
• Main Authors: Schreurs, Richard H. M. M, Legler, Juliette, Artola-Garicano, Elsa, Sinnige, Theo L, Lanser, Peter H, Seinen, Willem, van der Burg, Bart
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 15-02-2004
• Subjects: Agnatha. Pisces; Animal, plant and microbial ecology; Animals; Animals, Genetically Modified; Applied ecology; Benzopyrans - pharmacokinetics; Benzopyrans - toxicity; Biological and medical sciences; Body Weight; Cell Culture Techniques; Danio rerio; Dose-Response Relationship, Drug; Ecotoxicology, biological effects of pollution; Effects of pollution and side effects of pesticides on vertebrates; Endocrine System - drug effects; Estrogen Receptor Modulators - toxicity; Estrogens; Female; Fish; Freshwater; Fundamental and applied biological sciences. Psychology; Male; Perfume; Perfumes; Pollutants; Receptors, Estrogen - drug effects; Tetrahydronaphthalenes - pharmacokinetics; Tetrahydronaphthalenes - toxicity; Water Pollutants, Chemical - pharmacokinetics; Water Pollutants, Chemical - toxicity; Water treatment; Zebrafish; Zebrafish - physiology; Vertebrata; Musk; Pisces; Biota; Antiestrogen; Polycyclic compound; Water pollution; Cyprinidae; Biological effect; Biological activity; Contaminant
• ISSN: 0013-936X; 1520-5851
• DOI: 10.1021/es034648y

The polycyclic musks 6-acetyl-1,1,2,4,4,7-hexamethyltetraline (AHTN) and 1,2,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta-γ-2-benzopyran (HHCB) are used as fragrance ingredients in perfumes, soaps, and household cleaning products. They are known to be ubiquitously present in the aquatic environment, and because of their lipophilic nature, they tend to bioaccumulate in aquatic biota. In surface waters, concentrations between 1 ng/L and 5 μg/L have been found, depending mainly on the proportion of sewage effluents in the water. In fish, under normal environmental conditions, concentrations in the microgram per kilogram fresh weight (fw) range are found. In a previous study we showed that AHTN and HHCB exert mainly antiestrogenic effects on the human estrogen receptor α (ERα) and ERβ in an in vitro reporter gene assay. In the current study, we assessed the in vitro antiestrogenic effects of both musks on zebrafish ERs. Antagonism was observed on zfERβ, and more pronounced on the newly cloned zfERγ. Using a transgenic zebrafish assay, we studied antiestrogenicity of the musks in vivo. Dose-dependent antagonistic effects were observed at concentrations of 0.1 and 1 μM AHTN and HHCB. GC−MS analysis showed that the musks bioaccumulated in the fish, with internal concentrations (15−150 mg/kg fw) which were roughly 600 times higher than the nominal test doses. To our knowledge, this is the first time that environmental contaminants are shown to be antiestrogenic in an in vivo fish assay that focuses solely on ER-mediated effects. This makes the transgenic zebrafish assay a promising tool for the rapid detection of both estrogenic and antiestrogenic effects of chemicals in fish.