ISOLATION OF MICRONUCLEI FROM MOUSE-BLOOD AND FLUORESCENCE IN-SITU HYBRIDIZATION WITH A MOUSE CENTROMERIC DNA-PROBE
ISOLATION OF MICRONUCLEI FROM MOUSE-BLOOD AND FLUORESCENCE IN-SITU HYBRIDIZATION WITH A MOUSE CENTROMERIC DNA-PROBE
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DOI：
10.1016/0027-5107(94)90298-4
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发表时间：
1994-05-01
期刊：
影响因子：
--
通讯作者：
SOFUNI, T
中科院分区：
文献类型：
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作者：
HAYASHI, M;MAKIPAAKKANEN, J;SOFUNI, T
从小鼠血液中分离自发存在和化学诱导的微核。用PBS稀释50 μ l心脏血液并离心。此后，对细胞沉淀进行低渗处理，用乙酸-甲醇（1：3）固定，并通过2 μ m聚碳酸酯核孔膜过滤裂解物。将分离的微核在载玻片上风干，并使用小鼠着丝粒γ卫星探针进行荧光原位杂交（FISH）。从溶剂对照小鼠中分离的约一半微核显示出着丝粒信号。在这些初步研究中，纺锤体毒物（秋水仙碱和硫酸长春碱）处理增加了着丝粒阳性微核的比例，1-β-D-阿拉伯呋喃糖基胞嘧啶处理仅略微降低了着丝粒阳性微核的比例，并且通常不受丝裂霉素C的影响。
Spontaneously existing and chemically induced micronuclei were isolated from mouse blood. 50 mu l of cardiac blood was diluted with PBS and centrifuged. After this, the cell pellet was subjected to hypotonic treatment, fixed with acetic acid-methanol (1:3), and the lysate was filtrated through a 2-mu m polycarbonate nuclepore membrane. Isolated micronuclei were air-dried on a glass slide and subjected to fluorescence in situ hybridization (FISH) using a mouse centromeric gamma satellite probe. Approximately half of the micronuclei isolated from vehicle control mice showed centromere signal(s). In these preliminary studies, the proportion of centromere-positive micronuclei was increased by treatment with spindle poisons (colchicine and vinblastine sulfate), decreased only slightly by 1-beta-D-arabinofuranosylcytosine, and was generally unaffected by mitomycin C.