SIMULTANEOUS EVALUATION OF CLASTOGENICITY, ANEUGENICITY AND TOXICITY IN THE MOUSE MICRONUCLEUS ASSAY USING IMMUNOFLUORESCENCE
SIMULTANEOUS EVALUATION OF CLASTOGENICITY, ANEUGENICITY AND TOXICITY IN THE MOUSE MICRONUCLEUS ASSAY USING IMMUNOFLUORESCENCE
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DOI：
10.1016/0165-7992(92)90090-5
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发表时间：
1992-07-01
期刊：
影响因子：
--
通讯作者：
THEISS, JC
中科院分区：
文献类型：
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作者：
KRISHNA, G;FIEDLER, R;THEISS, JC
在小鼠微核实验中建立了一种改进的抗着丝粒抗体技术，可同时评价各种试验剂的毒性、致裂性和非致生性。该程序涉及使用纤维素柱分离细胞丝玻片进行分析。染色方法是用冠状血清、含氟的山羊抗人抗体和猪抗山羊抗体和碘化丙啶对载玻片进行顺序处理。在该方法中，使用相同的滤光片设置，在蓝色激发（440-490 nm）下（520 nm），识别多色红细胞（pce，暗红色）、正色红细胞（nce，绿色）、染色体/片段/微核（橙色）和着丝点（黄色）。采用该方法检测了环磷酰胺、x射线和长春新碱三种药物的微核/非整倍体诱导和骨髓毒性。异氰胺、长春新碱、碎裂原、x射线和环磷酰胺分别诱导着丝点阳性（K +）和阴性（K -）微核pce。在测试的剂量下，环磷酰胺引起雌性pce轻微但统计学上显著的下降，其他药物对雄性或雌性小鼠都没有产生任何严重的骨髓毒性。这些结果与文献中用各种方法报道的这些化合物的结果相比较，从而证明了该试验在区分致裂性和非致生性以及评估毒性方面的有效性。
An improved antikinetochore antibody technique was established in the mouse micronucleus assay to simultaneously evaluate toxicity, clastogenicity and aneugenicity induced by various test agents. The procedure involved the use of cellulose column fractionated cytospun slides for analysis. The staining method consisted of sequential treatment of slides with crest serum, fluorosceinated goat-antihuman and swine-antigoat antibodies, and propidium iodide. In this method, polychromatic erythrocytes (PCEs, dark red), normochromatic erythrocytes (NCEs, green), chromosome(s)/fragments/micronuclei (orange), and kinetochores (yellow), are identified using the same filter setting under blue excitation (440-490 nm) with a barrier filter at 520 nm. Using this method, three agents, cyclophosphamide, X-rays and vincristine were tested for micronucleus/ aneuploidy induction and bone marrow toxicity. The aneugen, vincristine, and clastogens, X-rays and cyclophosphamide, induced predominantly kinetochore positive (K + ) and negative (K - ) micronucleated PCEs, respectively. At the doses tested, cyclophosphamide caused a slight but statistically significant decrease in PCEs in females, and other agents did not produce any severe bone-marrow toxicity in either male or female mice. These results are comparable with the results reported in the literature on these compounds with various methods and thus demonstrate the usefulness of this assay in distinguishing clastogenicity from aneugenicity and in evaluating toxicity.