PUBLICATION
Development of a new screening assay to identify proteratogenic substances using zebrafish Danio rerio embryo combined with an exogenous mammalian metabolic activation system (mDarT)
- Authors
- Busquet, F., Nagel, R., von Landenberg, F., Mueller, S.O., Huebler, N., and Broschard, T.H.
- ID
- ZDB-PUB-080408-4
- Date
- 2008
- Source
- Toxicological sciences : an official journal of the Society of Toxicology 104(1): 177-188 (Journal)
- Registered Authors
- Nagel, Roland
- Keywords
- zebrafish, cyclophosphamide, ethanol, alternatives to animal testing, teratogenicity, metabolic activation
- MeSH Terms
-
- Animals
- Cyclophosphamide/toxicity*
- Embryo, Nonmammalian/abnormalities
- Embryo, Nonmammalian/drug effects
- Ethanol/toxicity*
- Microsomes, Liver/metabolism
- Rats
- Teratogens/toxicity*
- Toxicity Tests/methods*
- Zebrafish/abnormalities*
- PubMed
- 18375544 Full text @ Toxicol. Sci.
Citation
Busquet, F., Nagel, R., von Landenberg, F., Mueller, S.O., Huebler, N., and Broschard, T.H. (2008) Development of a new screening assay to identify proteratogenic substances using zebrafish Danio rerio embryo combined with an exogenous mammalian metabolic activation system (mDarT). Toxicological sciences : an official journal of the Society of Toxicology. 104(1):177-188.
Abstract
The assessment of teratogenic effects of chemicals is generally performed using in vivo teratogenicity assays e.g., in rats or rabbits. We have developed an in vitro teratogenicity assay using the zebrafish Danio rerio embryo combined with an exogenous mammalian metabolic activation system (MAS), able to biotransform proteratogenic compounds. Cyclophosphamide (CPA) and ethanol were used as proteratogens to test the efficiency of this assay. Briefly, the zebrafish embryos were co-cultured at 2 hpf (hours post fertilization) with the test material at varying concentrations, induced male rat liver microsomes and NADPH for 60 min at 32 degrees C under moderate agitation in Tris-buffesay. Briefly, the zebrafish embryos were co-cultured at 2 hpf (hours post fertilization) with the test material at varying concentrations, inr. The negative control (test material alone) and the MAS control (MAS alone) were incubated in parallel. For each test group, 20 eggs were used for statistical robustness. Afterwards fish embryos were transferred individually into 24-well plates filled with fish medium for 48 hours at 26 degrees C with a 12 hour-light cycle. Teratogenicity was scored after 24 and 48 hpf using morphological endpoints. No teratogenic effects were observed in fish embryos exposed to the proteratogens alone, i.e. without metabolic activation. In contrast, CPA and ethanol induced abnormalities in fish embryos when co-incubated with microsomes. The severity of malformations increased with increasing concentrations of the proteratogens. We conclude that the application of microsomes will improve and refine the Danio rerio teratogenicity assay as a predictive and valuable alternative method to screen teratogenic substances.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping