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³²P-Postlabeling Analysis of DNA Adducts in Liver of Wild English Sole (Parophrys vetulus) and Winter Flounder (Pseudopleuronectes americanus)¹

Environmental Conservation Division, Northwest and Alaska Fisheries Center, National Marine Fisheries Service, National Oceanic and Atmospheric Administration, Seattle, Washington 98112

The 1-butanol adduct enhancement version of the ³²P-postlabeling assay was used to measure the levels of hepatic DNA adducts in the marine flatfish, English sole (Parophrys vetulus), sampled from the Duwamish Waterway and Eagle Harbor, Puget Sound, WA, where they are exposed to high concentrations of sediment-associated chemical contaminants and exhibit an elevated prevalence of hepatic neoplasms. Hepatic DNA was also analyzed from English sole from a reference area (Useless Bay, WA) and from reference English sole treated with organicsolvent extracts of sediments from the two contaminated sites. Autoradiograms of thin-layer chromatograms of ³²P-labeled hepatic DNA digests from English sole from the contaminated sites exhibited up to three diagonal radioactive zones, which were not present in autoradiograms of thin-layer chromatogram maps of ³²P-labeled DNA digests from English sole from the reference site. These diagonal radioactive zones contained several distinct spots as well as what appeared to be multiple overlapping adduct spots. The levels (nmol of adducts/mol of nucleotides) of total DNA adducts for English sole from Duwamish Waterway and Eagle Harbor were 26 ± 28 (SD) and 17 ± 9.6, respectively. All autoradiograms of DNA from fish from the contaminated sites exhibited a diagonal radioactive zone where DNA adducts of chrysene, benzo(a)pyrene, and dibenz(a,h)anthracene, formed in vitro using English sole hepatic microsomes, were shown to chromatograph. English sole treated with extracts of the contaminated sediments had adduct profiles generally similar to those for English sole from the respective contaminated sites. The chromatographic characteristics of the adducts and the similarities in adduct profiles between field-sampled English sole and those treated with contaminated sediment extracts suggested that hydrophobic aromatic compounds of anthropogenic origin were adducted to hepatic DNA of sole from contaminated sites, but not in sole from the reference site. Moreover, an initial study with winter flounder (Pseudopleuronectes americanus) from Boston Harbor, MA, an area where high concentrations of sediment-associated chemicals are present, revealed a pattern of hepatic DNA-adducts (9.0 ± 7.8 nmol of adducts/mol of nucleotides) similar to that observed for English sole from Eagle Harbor.

¹ This project has been funded, at least in part, by the Office of Oceanography and Marine Assessments, NOAA. This work was presented in part at the 78th Annual Meeting of the American Association of Cancer Research, Atlanta, GA, 1987.

² To whom requests for reprints should be addressed, at Environmental Conservation Division Northwest and Alaska Fisheries Center, National Marine Fisheries Service, National Oceanic and Atmospheric Administration, 2725 Montlake Blvd., East, Seattle, WA 98112.

Received 7/ 1/88. Revised 10/17/88. Accepted 11/30/88.

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