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The Effects of the Cotreatments of Benzo(a)pyrene and Fluoranthene on the Expression and Activity of Cytochrome P450 1A and on the Formation of DNA Adducts in the Killifish, Fundulus heteroclitus

EPA Grant Number: U915631
Title: The Effects of the Cotreatments of Benzo(a)pyrene and Fluoranthene on the Expression and Activity of Cytochrome P450 1A and on the Formation of DNA Adducts in the Killifish, Fundulus heteroclitus
Investigators: Wassenberg, Deena M.
Institution: Duke University
EPA Project Officer: Bishop, Linda
Project Period: August 1, 1999 through August 1, 2002
Project Amount: $102,000
RFA: STAR Graduate Fellowships (1999)
Research Category: Fellowship - Life Sciences , Biology/Life Sciences , Academic Fellowships

Description:

Objective:

The objective of this research project is to find the effects of polynuclear aromatic hydrocarbons (PAHs), specifically benzo(a)pyrene (BaP) and fluoranthene (FL), alone and in combination, on the activity and expression of cytochrome P450 1A (CYP1A) and on the formation of deoxyribonucleic acid (DNA) adducts in the killifish, Fundulus heteroclitus.

Approach:

To explore the effects of mixed-contaminant exposures in vivo, F. heteroclitus will be dosed intraperitoneally with 5 mg/kg BaP, 5 mg/kg BaP + 5 mg/kg FL, 5 mg/kg BaP + 50 mg/kg FL, 5 mg/kg FL, 50 mg/kg FL, or corn oil. CYP1A mediated ethoxyresorufin-o-deethylase (EROD) activities of liver microsomes from each treatment group will be measured. Additionally, BaP induced microsomes will be incubated with 2 x 10-5 to 2 x 10-4 moles per liter (M) FL and tested for EROD activity. CYP1A protein expression of liver microsomes will be measured by Western blot, and PAH-DNA adduct formation will be analyzed for each treatment group by two dimensional thin-layer chromatography of 32P postlabeled nucleotides.

Supplemental Keywords:

Fundulus heteroclitus, killifish, polynuclear aromatic hydrocarbons, PAHs, benzo(a)pyrene, BaP, fluoranthene, FL, CYP1A, biomarker, deoxyribonucleic acid, DNA, DNA adducts, ethoxyresorufin-o-deethylase, EROD.

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The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.

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