Molecular multi-effect screening of environmental pollutants using the MolDarT.
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 (mt2), cytochrome P450 1A1 (cyp1a1), and recombination activation gene 1 (rag1) are used as endpoints that describe detoxification/metal toxicity (mt2), detoxification/PAH toxicity (cyp1a1), and acquired immune system disruption (rag1). 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, mt2 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 microM ZnSO(4) significantly induced mt2; 10 microM DMN and 20 microM DMN resulted in significantly increased cyp1a1 abundance; and 5 and 10 microM CdCl(2) significantly reduced rag1 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 microM) significantly lowered mt2 expression. This study shows the potential of short-term in vivo multi-effect screenings within one single subacute exposure using the MolDarT.
Liedtke
Anja
A
Eawag, Swiss Federal Institute of Aquatic Science and Technology, Environmental Toxicology, Ueberlandstrasse 133, 8600, Dübendorf, Switzerland.
Muncke
Jane
J
Rüfenacht
Karin
K
Eggen
Rik I L
RI
D000818:Animals; D019256:Cadmium Chloride / Q000633:toxicity; D019363:Cytochrome P-450 CYP1A1 / Q000187:drug effects / Q000235:genetics; D017931:DNA Primers; D004781:Environmental Exposure; D004784:Environmental Monitoring / Q000379:methods; D004785:Environmental Pollutants / Q000633:toxicity*; D004967:Estrogens / Q000633:toxicity*; D015870:Gene Expression / Q000187:drug effects; D009281:Naphthalenes / Q000633:toxicity; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015027:Zebrafish / Q000235:genetics*; D029961:Zebrafish Proteins / Q000187:drug effects* / Q000235:genetics; D019287:Zinc Sulfate / Q000633:toxicity
D017931:DNA Primers; D004785:Environmental Pollutants; D004967:Estrogens; D009281:Naphthalenes; D012333:RNA, Messenger; D029961:Zebrafish Proteins; D019287:Zinc Sulfate; D019363:Cytochrome P-450 CYP1A1; D019256:Cadmium Chloride