Suzuki T, Tamai K, Kodama Y, Asita A O, Matsuoka A, Sofuni T, Kurita M, Ohtsuki H, Hiwatashi T, Hayashi M
Division of Genetics and Mutagenesis, National Institute of Hygiene Sciences, Tokyo, Japan.
Mutat Res. 1992 Feb-Mar;278(2-3):169-73. doi: 10.1016/0165-1218(92)90229-s.
Micronucleus assays using mouse peripheral blood stained vitally on acridine orange (AO)-coated slides were evaluated at two laboratories with 7,12-dimethylbenz[a]anthracene (DMBA) and compared with the standard bone marrow assay. DMBA was administered by single intraperitoneal injection to CD-1 mice at doses ranging from 5 to 80 mg/kg, then 5 microliters of peripheral blood was sampled from a tail vein at 24, 48, 72, 96, and 120 h after treatment. Similar incidences of micronucleated young erythrocytes were observed in peripheral blood reticulocytes and bone marrow polychromatic erythrocytes. The dose response of micronucleated reticulocytes was delayed compared to that of micronucleated polychromatic erythrocytes. The dose-response curves after treatment with DMBA differed depending on the sampling times, which revealed the difficulty of obtaining accurate dose-response relations in the micronucleus assay. The present result demonstrated that the simple and rapid AO supravital staining method is a valuable and easier method for obtaining dose- and time-response data for quantification of micronucleus induction by chemicals.
在两个实验室中，对使用吖啶橙（AO）包被玻片进行活体染色的小鼠外周血进行微核试验，以评估7,12-二甲基苯并[a]蒽（DMBA），并与标准骨髓试验进行比较。将DMBA以5至80 mg/kg的剂量通过单次腹腔注射给予CD-1小鼠，然后在治疗后24、48、72、96和120小时从尾静脉采集5微升外周血。在外周血网织红细胞和骨髓多染性红细胞中观察到微核幼红细胞的发生率相似。与微核多染性红细胞相比，微核网织红细胞的剂量反应有所延迟。用DMBA处理后的剂量反应曲线因采样时间而异，这表明在微核试验中获得准确的剂量反应关系存在困难。目前的结果表明，简单快速的AO活体染色方法是一种有价值且更简便的方法，可用于获取化学物质诱导微核定量的剂量和时间反应数据。
