Detection of rodent liver carcinogen genotoxicity by the alkaline single-cell gel electrophoresis (Comet) assay in multiple mouse organs (liver, lung, spleen, kidney, and bone marrow)

• Published in: Mutation research Vol. 391; no. 3; pp. 201 - 214
• Main Authors: Sasaki, Yū F, Izumiyama, Fusako, Nishidate, Emi, Matsusaka, Naonori, Tsuda, Shuji
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 14-07-1997
• Subjects: Alkaline single cell gel electrophoresis (SCG) assay; Animals; Benzophenoneidum - toxicity; Bone Marrow - drug effects; Carcinogens - toxicity; Chlorobenzenes - toxicity; Comet assay; DNA lesion; Electrophoresis - methods; Epoxy Compounds - toxicity; Ethylenethiourea - toxicity; Hexachlorocyclohexane - toxicity; Kidney - drug effects; Liver; Liver - drug effects; Liver carcinogen; Lung - drug effects; Male; Mice; Mice, Inbred Strains; Micronucleus Tests; Mouse; Multiple organ; Mutagenicity Tests - methods; Organ Specificity; p-Aminoazobenzene - toxicity; Phenobarbital - toxicity; Phenylenediamines - toxicity; Spleen - drug effects; Alkaline single cell gel electrophoresis (SCG) assay; Liver carcinogen; Mouse; Liver; DNA lesion; Multiple organ; Comet assay
• ISSN: 1383-5718; 0027-5107; 1879-3592
• DOI: 10.1016/S1383-5718(97)00072-7

We have recently designed a simple method for applying the alkaline single-cell gel electrophoresis (SCG) assay to mouse organs. With this method, each organ is minced, suspended in chilled homogenizing buffer containing NaCl and Na
2EDTA, gently homogenized using a Potter-type homogenizer set in ice, and then centrifuged nuclei are used for the alkaline SCG assay. In the present study, we used the method to assess the genotoxicity of 8 rodent hepatic carcinogens in 5 mouse organs (liver, lung, kidney, spleen, and bone marrow). The carcinogens we studied were
p-aminoazobenzene, auramine, 2,4-diaminotoluene,
p-dichlorobenzene, ethylene thiourea (ETU), styrene-7,8-oxide, phenobarbital sodium, and benzene-1,2,3,4,5,6-hexachloride (BHC); except for
p-aminoazobenzene, they do not induce micronuclei in mouse bone marrow cells. Mice were sacrificed 3 and 24 h after the administration of each carcinogen.
p-Aminoazobenzene, ETU, and styrene-7,8-oxide induced alkaline labile DNA lesions in all of the organs studied. Auramine, 2,4-diaminotoluene,
p-dichlorobenzene, and phenobarbital sodium also produced lesions, but their effect was greatest in the liver. BHC, which is not genotoxic in in vitro tests, did not show any effects. We suggest that it may be possible to use the alkaline SCG assay to detect in vivo activity of chemicals whose genotoxicity is not expressed in bone marrow cells.