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Title: Detection of genotoxic insult as DNA strand breaks in fish blood cells by agarose gel electrophoresis

Abstract

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.

Authors:
 [1];  [2];  [3]
  1. Univ. of Tennessee, Knoxville, TN (United States)
  2. Univ. of Mississippi, University, MS (United States). Dept. of Biology
  3. Oak Ridge National Lab., TN (United States). Environmental Sciences Division
Publication Date:
OSTI Identifier:
7017038
Resource Type:
Journal Article
Journal Name:
Environmental Toxicology and Chemistry; (United States)
Additional Journal Information:
Journal Volume: 13:7; Journal ID: ISSN 0730-7268
Country of Publication:
United States
Language:
English
Subject:
54 ENVIRONMENTAL SCIENCES; 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

Citation Formats

Theodorakis, C W, D'Surney, S J, and Shugart, L R. Detection of genotoxic insult as DNA strand breaks in fish blood cells by agarose gel electrophoresis. United States: N. p., 1994. Web. doi:10.1002/etc.5620130703.
Theodorakis, C W, D'Surney, S J, & Shugart, L R. Detection of genotoxic insult as DNA strand breaks in fish blood cells by agarose gel electrophoresis. United States. https://doi.org/10.1002/etc.5620130703
Theodorakis, C W, D'Surney, S J, and Shugart, L R. 1994. "Detection of genotoxic insult as DNA strand breaks in fish blood cells by agarose gel electrophoresis". United States. https://doi.org/10.1002/etc.5620130703.
@article{osti_7017038,
title = {Detection of genotoxic insult as DNA strand breaks in fish blood cells by agarose gel electrophoresis},
author = {Theodorakis, C W and D'Surney, S J and Shugart, L R},
abstractNote = {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.},
doi = {10.1002/etc.5620130703},
url = {https://www.osti.gov/biblio/7017038}, journal = {Environmental Toxicology and Chemistry; (United States)},
issn = {0730-7268},
number = ,
volume = 13:7,
place = {United States},
year = {Fri Jul 01 00:00:00 EDT 1994},
month = {Fri Jul 01 00:00:00 EDT 1994}
}