Alkaline elution of rat testicular DNA: detection of DNA cross-links after in vivo treatment with chemical mutagens.
The alkaline elution technique was used to measure DNA damage in the rat testis after intraperitoneal injection of 3 chemicals known to cause heritable mutations in rodents. These 3 chemicals are triethylenemelamine (TEM), mitomycin C, and cyclophosphamide. All three of these chemicals produced DNA damage which was readily detectable by alkaline elution. Both TEM and mitomycin C produced DNA interstrand cross-links, although TEM was a more potent cross-linker on an equimolar basis than mitomycin C. Cyclophosphamide produced both DNA cross-links and DNA strand breaks. Alkaline elution in the absence of proteinase K indicated that some of the strand breaks appeared to be closely associated with protein. These studied indicate that the alkaline elution technique is capable of detecting DNA damage in mammalian germ cells produced by chemical mutagens. This technique may prove useful as a screening tool for identifying chemicals which cause heritable mutations in mammals.
D000818:Animals; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D003520:Cyclophosphamide / Q000494:pharmacology; D004247:DNA / Q000032:analysis*; D019286:Endopeptidase K; D010450:Endopeptidases; D006863:Hydrogen-Ion Concentration; D008297:Male; D008937:Mitomycins / Q000494:pharmacology; D009152:Mutagenicity Tests; D009153:Mutagens / Q000494:pharmacology*; D051381:Rats; D013737:Testis / Q000032:analysis*; D014265:Triethylenemelamine / Q000494:pharmacology
D003432:Cross-Linking Reagents; D008937:Mitomycins; D009153:Mutagens; D003520:Cyclophosphamide; D004247:DNA; D010450:Endopeptidases; D019286:Endopeptidase K; D014265:Triethylenemelamine