Detection of chemically induced DNA lesions in multiple-mouse organs (liver, lung, spleen, kidney, and bone marrow) using the alkaline single cell gel electrophoresis (Comet) assay
Detection of chemically induced DNA lesions in multiple-mouse organs (liver, lung, spleen, kidney, and bone marrow) using the alkaline single cell gel electrophoresis (Comet) assay
复制标题
DOI：
10.1016/s1383-5718(96)00133-7
复制
发表时间：
1997-01-15
影响因子：
1.9
通讯作者：
Nishidate, E
中科院分区：
文献类型：
--
作者：
Sasaki, YF;Tsuda, S;Nishidate, E
用碱性单细胞凝胶电泳（SCG）（彗星试验）研究了2种典型化学诱变剂对小鼠肝、肺、肾、脾和骨髓5个器官DNA的影响。在给予直接诱变剂乙基亚硝基脲（ENU）或肝靶向前诱变剂对二甲氨基偶氮苯（DAB）后3和24 h处死小鼠。将每个器官切碎，以1 μ g/ml的浓度悬浮于含有0.075 M NaCl和0.024 M Na（2）EDTA的冷冻的Na 2SO 4缓冲液（pH 7.5）中，使用Potter型均质器在冰上以500-800 rpm的转速轻轻均质化，然后将离心的细胞核用于碱性SCG测定。ENU在所研究的所有器官的细胞中诱导DNA损伤，另一方面，DAB仅在肝脏中产生阳性反应。我们建议，它可能是可能的，使用碱性SCG试验，采用匀浆技术，以检测在体内的化学品在其靶器官的遗传毒性。
The effect of 2 model chemical mutagens on DNA was evaluated with the alkaline single cell gel electrophoresis (SCG) (Comet) assay in 5 mouse organs - liver, lung, kidney, spleen and bone marrow. Mice were sacrificed 3 and 24 h after the administration of the direct mutagen ethyl nitrosourea (ENU) or the liver-targeting promutagen p-dimethylaminoazobenzene (DAB). Each organ was minced, suspended at a concentration of 1 g/ml in chilled homogenizing buffer (pH 7.5) containing 0.075 M NaCl and 0.024 M Na(2)EDTA, homogenized gently using a Potter-type homogenizer at 500-800 rpm set in ice, and then centrifuged nuclei were used for the alkaline SCG assay. ENU induced DNA damage in cells all of the organs studied, DAB, on the other hand, produced a positive response in the liver only. We suggest that it may be possible to use the alkaline SCG assay using a homogenization technique to detect the genotoxicity of chemicals in vivo in their target organs.