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Molecular multi‐effect screening of environmental pollutants using the MolDarT
Author(s) -
Liedtke Anja,
Muncke Jane,
Rüfenacht Karin,
Eggen Rik I. L.
Publication year - 2008
Publication title -
environmental toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.813
H-Index - 77
eISSN - 1522-7278
pISSN - 1520-4081
DOI - 10.1002/tox.20305
Subject(s) - zebrafish , danio , nonylphenol , toxicity , gene expression , cytochrome p450 , gene , environmental toxicology , chemistry , biology , bisphenol a , pollutant , microbiology and biotechnology , environmental chemistry , biochemistry , enzyme , ecology , organic chemistry , epoxy
Molecular effect detection is a useful approach for ecotoxicological screening of chemicals. We show here the application of the molecular DarT (MolDarT), where the expression of selected target genes is detected in short‐term (120 h) exposed developing zebrafish ( Danio rerio ), thus allowing subacute multi‐effect compound screening. The genes metallothionein 2 ( mt 2 ), cytochrome P450 1A1 ( cyp 1a 1 ), and recombination activation gene 1 ( rag 1 ) are used as endpoints that describe detoxification/metal toxicity ( mt 2 ), detoxification/PAH toxicity ( cyp 1a 1 ), and acquired immune system disruption ( rag 1 ). Each gene's developmental expression was studied in unexposed zebrafish during 4 to 120 h past fertilization (hpf), and all three genes were found to be expressed at 120 hpf. Furthermore, mt 2 transcripts were present at high levels at 4 hpf, indicating a maternal transfer. For positive toxicity controls, freshly fertilized zebrafish eggs were exposed for 120 hpf to ZnSO 4 , 1,5‐dimethylnaphthalene (DMN) and CdCl 2 . Exposure to 100 and 200 μM ZnSO 4 significantly induced mt 2 ; 10 μM DMN and 20 μM DMN resulted in significantly increased cyp 1a 1 abundance; and 5 and 10 μM CdCl 2 significantly reduced rag 1 expression levels. Furthermore, we analysed these target genes for their expression in zebrafish eggs from a previous exposure study. The eggs were exposed for 120 hpf to the environmental pollutants estradiol (E2), ethinylestradiol (EE2), nonylphenol (NP), atrazine, cyproconazol, and bisphenol A (BPA) and found differential expression of the three genes. Exposure to the (xeno‐)estrogenic compound NP (0.75 μM) significantly lowered mt 2 expression. This study shows the potential of short‐term in vivo multi‐effect screenings within one single subacute exposure using the MolDarT. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2008.