Premium
Non destructive in vivo measurement of ethoxyresorufin biotransformation by zebrafish prolarva: Development and application
Author(s) -
Noury P.,
Geffard O.,
Tutundjian R.,
Garric J.
Publication year - 2006
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.20184
Subject(s) - danio , biotransformation , bioassay , zebrafish , chemistry , cytochrome p450 , environmental chemistry , xenobiotic , in vivo , biochemistry , biology , toxicology , chromatography , enzyme , microbiology and biotechnology , ecology , gene
Biotransformation enzymatic activities, such as the cytochrome P450 one, have been used as biomarkers for pollution assessment for a long time. Nevertheless, such biochemical tools are destructive processes, because they are performed on fish liver or total larvae homogenates. Moreover, the adaptation of this bioassay to some fish larvae, like Danio rerio ones, is ineffective because it needs a lot of organisms. We thus developed an original, nondestructive method to detect the induction of a biotransformation activity in the prolarva of the fish, Danio rerio , exposed to chemicals. This methodology is based on the assessment of EROD activity, by measurement in the culture medium of the fluorescence increase due to the excretion of resorufin by fish during an ethoxyresorufin exposure. After exposure of fish embryos to known inducers (B a P and β‐naphtoflavone), the prolarvae were exposed to the substrate (ethoxyresorufin), and the kinetic of the fluorescence increase was measured. A dose–effect relationship was observed, with a three to five fold increase of EROD basal activity. This methodology also allowed us to compare between EROD activity induction by different environmental samples. The proposed methodology thus allows to perform a simple, sensitive, and reproducible microbiotest for the detection of sublethal concentrations of AhR chemical inducers in environmental samples. © 2006 Wiley Periodicals, Inc. Environ Toxicol 21: 324–331, 2006.