Detection of genotoxic insult as DNA strand breaks in fish blood cells by agarose gel electrophoresis
- Univ. of Tennessee, Knoxville, TN (United States)
- Univ. of Mississippi, University, MS (United States). Dept. of Biology
- Oak Ridge National Lab., TN (United States). Environmental Sciences Division
DNA, isolated from the blood cells of bluegill sunfish (Lepomis macrochirus) exposed in the lab to bedded sediment collected from a site contaminated with genotoxic compounds (i.e., PAHs, PCBs, and heavy metals), was examined for strand breakage by agarose gel electrophoresis. Before electrophoresis the blood cells were embedded in agarose plugs and incubated with proteinase. After electrophoresis under both neutral (pH 7) or alkaline (pH 12) conditions, the median molecular length (MML) of the DNA distributed in the gel was determined. These quantitative measures were used to estimate the difference in the number of double- and single-strand breaks between DNA preparations. Both types of strand breakage were found to be greater in fish exposed to sediment contaminated with genotoxic compounds as compared to nonexposed fish. A statistically significant correlation was demonstrated between the MML value obtained by the electrophoretic assay reported here and the F value (measure of DNA double-strandedness) obtained by the alkaline unwinding assay.
- OSTI ID:
- 7017038
- Journal Information:
- Environmental Toxicology and Chemistry; (United States), Vol. 13:7; ISSN 0730-7268
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
FISHES
SENSITIVITY
METALS
GENETIC EFFECTS
POLYCHLORINATED BIPHENYLS
POLYCYCLIC AROMATIC HYDROCARBONS
SEDIMENTS
CONTAMINATION
STRAND BREAKS
ELECTROPHORESIS
WATER POLLUTION
ANIMALS
AQUATIC ORGANISMS
AROMATICS
BIOLOGICAL EFFECTS
CHLORINATED AROMATIC HYDROCARBONS
ELEMENTS
HALOGENATED AROMATIC HYDROCARBONS
HYDROCARBONS
ORGANIC CHLORINE COMPOUNDS
ORGANIC COMPOUNDS
ORGANIC HALOGEN COMPOUNDS
POLLUTION
VERTEBRATES
540320* - Environment
Aquatic- Chemicals Monitoring & Transport- (1990-)
560300 - Chemicals Metabolism & Toxicology