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Improved single-cell gel electrophoresis assay for detecting DNA damage in Eisenia foetida.
Environ Mol Mutagen. 2005 Dec; 46(4):246-52.EM

Abstract

The earthworm (Eisenia foetida) is an attractive sentinel species for detecting genotoxicity in soil. In this study, an improved single-cell gel electrophoresis (SCGE) assay was developed for detecting DNA damage in the coelomocytes (lymphocytes) of earthworms. Coelomocytes were obtained from the coelomic fluid using a modified extrusion medium that did not include the mucolytic agent guaiacol. The extruded coelomocytes contained at least three types of cells: eleocytes (75% of the total), amoebocytes, and granulocytes. The DNA migration parameters were determined for untreated cells of each type in order that the assay could be performed with minimum inter- and intra-individual variation. In addition, lysis time was reduced to 10 min, and only one neutralization step was used. DNA damage was detected in isolated eleocytes treated with hydrogen peroxide and cadmium, and in eleocytes from earthworms exposed for up to 21 days to soil containing polycyclic aromatic hydrocarbons. The SCGE assay using earthworm eleocytes appears to be a sensitive biomarker for evaluating exposure to genotoxic compounds.

Authors+Show Affiliations

Programa de Investigación en Ecotoxicología, Departamento de Ciencias Básicas, Universidad Nacional de Luján, Rutas 5 y 7 s/n, 6700 Luján, Argentina. dimarzio@speedy.com.arNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15957191

Citation

Di Marzio, Walter D., et al. "Improved Single-cell Gel Electrophoresis Assay for Detecting DNA Damage in Eisenia Foetida." Environmental and Molecular Mutagenesis, vol. 46, no. 4, 2005, pp. 246-52.
Di Marzio WD, Saenz ME, Lemière S, et al. Improved single-cell gel electrophoresis assay for detecting DNA damage in Eisenia foetida. Environ Mol Mutagen. 2005;46(4):246-52.
Di Marzio, W. D., Saenz, M. E., Lemière, S., & Vasseur, P. (2005). Improved single-cell gel electrophoresis assay for detecting DNA damage in Eisenia foetida. Environmental and Molecular Mutagenesis, 46(4), 246-52.
Di Marzio WD, et al. Improved Single-cell Gel Electrophoresis Assay for Detecting DNA Damage in Eisenia Foetida. Environ Mol Mutagen. 2005;46(4):246-52. PubMed PMID: 15957191.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Improved single-cell gel electrophoresis assay for detecting DNA damage in Eisenia foetida. AU - Di Marzio,Walter D, AU - Saenz,María E, AU - Lemière,Sebastien, AU - Vasseur,Paule, PY - 2005/6/16/pubmed PY - 2006/3/18/medline PY - 2005/6/16/entrez SP - 246 EP - 52 JF - Environmental and molecular mutagenesis JO - Environ Mol Mutagen VL - 46 IS - 4 N2 - The earthworm (Eisenia foetida) is an attractive sentinel species for detecting genotoxicity in soil. In this study, an improved single-cell gel electrophoresis (SCGE) assay was developed for detecting DNA damage in the coelomocytes (lymphocytes) of earthworms. Coelomocytes were obtained from the coelomic fluid using a modified extrusion medium that did not include the mucolytic agent guaiacol. The extruded coelomocytes contained at least three types of cells: eleocytes (75% of the total), amoebocytes, and granulocytes. The DNA migration parameters were determined for untreated cells of each type in order that the assay could be performed with minimum inter- and intra-individual variation. In addition, lysis time was reduced to 10 min, and only one neutralization step was used. DNA damage was detected in isolated eleocytes treated with hydrogen peroxide and cadmium, and in eleocytes from earthworms exposed for up to 21 days to soil containing polycyclic aromatic hydrocarbons. The SCGE assay using earthworm eleocytes appears to be a sensitive biomarker for evaluating exposure to genotoxic compounds. SN - 0893-6692 UR - https://www.unboundmedicine.com/medline/citation/15957191/Improved_single_cell_gel_electrophoresis_assay_for_detecting_DNA_damage_in_Eisenia_foetida_ L2 - https://doi.org/10.1002/em.20153 DB - PRIME DP - Unbound Medicine ER -