Kinetics of mixed function oxygenase induction and retene excretion in retene-exposed rainbow trout (Oncorhynchus mykiss).

Abstract

The polycyclic aromatic hydrocarbon 7-isopropyl-1-methylphenanthrene (retene) induces mixed function oxygenase (MFO) activity of fish. Bile levels of retene and its metabolite(s) were measured in relation to exposure time, exposure concentration, and induction of MFO activity. Synchronous fluorescence spectrometry provided a rapid means of measuring the amount of retene present in the bile of exposed fish, whereas conventional fluorescence spectrometry was used to quantify the amount of retene metabolites. Based on bile analysis, increased retene exposure resulted in an increased uptake of retene and a curvilinear increase in hepatic MFO activity. Retene was present in the bile within 6 h of initial exposure. However, retene metabolite(s) only appeared in the bile after MFO induction had occurred, 12 h after exposure had commenced, suggesting that MFO activity is required for metabolism. Transfer of fish to clean water after 48 h of exposure resulted in a rapid decrease in the presence of retene and its metabolite(s) in the bile, with a calculated half-life of about 14 h. In vitro additions of retene directly to the ethoxyresorufin O-deethylase assay demonstrated that retene is capable of acting as a competitive inhibitor. Thus, retene contamination of postmitochondrial supernatant (S9 fraction) could result in false-negative results in the MFO assay. The MFO activity in extrahepatic tissues (gills, heart, and kidney) was not significantly induced with retene exposure. Thus, the major site of retene metabolism seems to be in the liver. These results confirm that retene is rapidly taken up, metabolized, and excreted by rainbow trout, and that retene metabolism and excretion are linked to hepatic MFO induction.