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).
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 Na2EDTA, 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.
Sasaki
Y F
YF
Laboratory of Genotoxicity, Faculty of Chemical and Biological Engineering, Hachinohe National College of Technology, Aomori, Japan. yfsasaki-c@hachinohe-ct.ac.jp
Izumiyama
F
F
Nishidate
E
E
Matsusaka
N
N
Tsuda
S
S
D000818:Animals; D001576:Benzophenoneidum / Q000633:toxicity; D001853:Bone Marrow / Q000187:drug effects; D002273:Carcinogens / Q000633:toxicity*; D002722:Chlorobenzenes / Q000633:toxicity; D004586:Electrophoresis / Q000379:methods*; D004852:Epoxy Compounds / Q000633:toxicity; D005031:Ethylenethiourea / Q000633:toxicity; D001556:Hexachlorocyclohexane / Q000633:toxicity; D007668:Kidney / Q000187:drug effects; D008099:Liver / Q000187:drug effects*; D008168:Lung / Q000187:drug effects; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D015162:Micronucleus Tests; D009152:Mutagenicity Tests / Q000379:methods*; D009928:Organ Specificity; D010634:Phenobarbital / Q000633:toxicity; D010655:Phenylenediamines / Q000633:toxicity; D013154:Spleen / Q000187:drug effects; D010128:p-Aminoazobenzene / Q000633:toxicity
D002273:Carcinogens; D002722:Chlorobenzenes; D004852:Epoxy Compounds; D010655:Phenylenediamines; D001576:Benzophenoneidum; D005031:Ethylenethiourea; D010128:p-Aminoazobenzene; D001556:Hexachlorocyclohexane; C013690:styrene oxide; C018511:4-dichlorobenzene; C010914:2,4-diaminotoluene; C023888:beta-hexachlorocyclohexane; D010634:Phenobarbital