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An in vitro rainbow trout cell bioassay for aryl hydrocarbon receptor‐mediated toxins
Author(s) -
Richter Catherine A.,
Tieber Virginia L.,
Denison Michael S.,
Giesy John P.
Publication year - 1997
Publication title -
environmental toxicology and chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.1
H-Index - 171
eISSN - 1552-8618
pISSN - 0730-7268
DOI - 10.1002/etc.5620160321
Subject(s) - bioassay , aryl hydrocarbon receptor , rainbow trout , toxicity , trout , luciferase , reporter gene , cell culture , environmental chemistry , chemistry , biology , ec50 , receptor , in vitro , transfection , biochemistry , pharmacology , fish <actinopterygii> , ecology , gene expression , gene , fishery , genetics , organic chemistry , transcription factor
Halogenated aromatic hydrocarbons (HAHs) and other chemicals that act as aryl hydrocarbon (Ah) receptor (AhR) agonists cause a variety of toxic effects. In sac fry of many fish species, these effects include blue‐sac disease and mortality. Because HAHs occur in complex mixtures, their toxicity in the environment is difficult to predict. A bioassay useful in predicting AhR‐mediated toxicity to fish was developed using the RTH‐149 rainbow trout hepatoma cell line. Stable transfection of this cell line with the pGudLuc1.1 plasmid, which contains a firefly luciferase reporter gene under the transcriptional regulation of dioxin responsive enhancers, has produced a recombinant cell line designated Remodulated Lightning Trout (RLT 2.0). The RLT 2.0 bioassay method detection limit for 2,3,7,8‐tetrachlorodibenzo‐ p ‐dioxin (TCDD) is 4 pM. The responses of the RLT 2.0 bioassay to TCDD and several HAH congeners closely matched the responses observed in vivo in fish. The RLT 2.0 bioassay can provide an integrative measure of the total AhR‐mediated toxic activity of complex mixtures to fish. The assay will be useful in screening environmental extracts, guiding chemical analysis, and interpreting the AhR‐mediated mechanism of toxicity.