ANALYSIS OF MICRONUCLEI INDUCED IN MOUSE EARLY SPERMATIDS BY MITOMYCIN-C, VINBLASTINE SULFATE OR ETOPOSIDE USING FLUORESCENCE IN-SITU HYBRIDIZATION

被引:43
作者
KALLIO, M
LAHDETIE, J
机构
[1] Department of Medical Genetics, University of Turku, SF-20520 Turku
基金
芬兰科学院;
关键词
D O I
10.1093/mutage/8.6.561
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
Non-radioactive in situ hybridization with mouse centromere specific (major) gamma satellite DNA probe was used to analyze the mechanism of induction of spermatid micronuclei (MN) caused by the alkylating agent mitomycin C (MMC), the spindle poison vinblastine sulfate (VBL) or the DNA topoisomerase II inhibitor etoposide (VP-16). Male mice were treated with a single i.p. injection of 25 mg/kg VP-16, 5 mg/kg MMC or 2 mg/kg VBL, respectively. After 24 h (VP-16, VBL) or 13 days (MMC) stage I spermatid slides were prepared and in situ hybridization was performed using a polymerase chain reaction amplified mouse (major) gamma satellite DNA probe. The observed MN frequencies for VP-16 and MMC, 6.2/1000 and 7.5/1000 round spermatids, respectively, show a strong mutagenic effect on mouse germ cells compared with controls (1.4/1000 spermatids). VBL, on the contrary, induced a much lower total frequency of MN (2.8/1000 spermatids) compared with previous results on mouse somatic cells. Of MN in controls, 24% carried a FISH signal. After correcting for background, MMC induced 38.6% signal-positive MN, consistent with a predominantly clastogenic mode of action, while VBL induced 67.9% signal-positive MN, consistent with a mainly aneugenic mechanism. VP-16 induced 65.5% signal-positive MN, indicating that its MN-inducing capacity is mainly due to whole chromosome lagging.
引用
收藏
页码:561 / 567
页数:7
相关论文
共 54 条
[21]   MEIOTIC MICRONUCLEI INDUCED BY X-RAYS IN EARLY SPERMATIDS OF THE RAT [J].
LAHDETIE, J ;
PARVINEN, M .
MUTATION RESEARCH, 1981, 81 (01) :103-115
[22]  
LAHDETIE J, 1982, PREVENTION OCCUPATIO, P483
[23]   HIGHER-ORDER METAPHASE CHROMOSOME STRUCTURE - EVIDENCE FOR METALLOPROTEIN INTERACTIONS [J].
LEWIS, CD ;
LAEMMLI, UK .
CELL, 1982, 29 (01) :171-181
[24]  
MA XJ, 1993, J BIOL CHEM, V268, P6182
[25]   THE SPIRAL STRUCTURE OF CHROMOSOMES [J].
MANTON, I .
BIOLOGICAL REVIEWS, 1950, 25 (04) :486-508
[26]   ACTION OF MITOMYCIN-C ON MOUSE SPERMATOGONIA [J].
MANYAK, A ;
SCHLEIERMACHER, E .
MUTATION RESEARCH, 1973, 19 (01) :99-108
[27]   ANALYSIS OF MICRONUCLEI INDUCED BY 2-CHLOROBENZYLIDENE MALONITRILE (CS) USING FLUORESCENCE INSITU HYBRIDIZATION WITH TELOMERIC AND CENTROMERIC DNA PROBES, AND FLOW-CYTOMETRY [J].
MILLER, BM ;
NUSSE, M .
MUTAGENESIS, 1993, 8 (01) :35-41
[28]   ANALYSIS OF RADIATION-INDUCED MICRONUCLEI BY FLUORESCENCE INSITU HYBRIDIZATION (FISH) SIMULTANEOUSLY USING TELOMERIC AND CENTROMERIC DNA PROBES [J].
MILLER, BM ;
WERNER, T ;
WEIER, HU ;
NUSSE, M .
RADIATION RESEARCH, 1992, 131 (02) :177-185
[29]   CLASSIFICATION OF MICRONUCLEI IN MURINE ERYTHROCYTES - IMMUNOFLUORESCENT STAINING USING CREST ANTIBODIES COMPARED TO INSITU HYBRIDIZATION WITH BIOTINYLATED GAMMA SATELLITE DNA [J].
MILLER, BM ;
ZITZELSBERGER, HF ;
WEIER, HUG ;
ADLER, ID .
MUTAGENESIS, 1991, 6 (04) :297-302
[30]  
MORAD M, 1973, HEREDITAS, V74, P273