Frameshift mutations induced by the acridine mustard ICR-191 in embryos and in the adult gill and hepatopancreas of rpsL transgenic zebrafish
Frameshift mutations induced by the acridine mustard ICR-191 in embryos and in the adult gill and hepatopancreas of rpsL transgenic zebrafish
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DOI：
10.1016/j.mrfmmm.2005.05.006
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发表时间：
2005-10-15
影响因子：
2.3
通讯作者：
Aoki, Y
中科院分区：
文献类型：
--
作者：
Nakamura, T;Amanuma, K;Aoki, Y
为了确定是否可以在rpsL转基因斑马鱼（斑马鱼）中检测到移码突变，用6-氯-9- [3-（2-氯乙氨基）-丙氨基] -2-甲氧基吖啶（ICR-191）处理胚胎和成鱼。在水浴中暴露于0、10或20 μ M ICR-191 18小时的胚胎显示出的诱导突变频率（MF）分别为14 x 10（-5）、16 x 10（-5）和25 x 10（-5）。只有暴露于20 μ M ICR- 191的胚胎显示ME显著增加。对照组和ICR-191处理组之间的突变谱不同，在10和20 μ M处理的胚胎中均观察到单个G：C对插入，这是ICR- 191诱变的显著特征。在水浴中用1 μ M ICR- 191处理成鱼18小时后，暴露2周后，在鳃（对照组和处理组鱼分别为12 x 10-5和44 x 10-5）和肝胰腺（分别为5 x 10 - 5和29 x 10 - 5）中观察到MF显著增加。序列分析表明，鳃中58%的突变和肝胰腺中94%的突变是单个G：C对插入，这是ICR-191诱导的典型突变。此外，这些突变主要发生在rpsL基因中的单个位点（CC序列在bp 140-141处）。然而，暴露后三周，ICR处理的鱼的增加的MF和突出的突变谱是检测不到的。这些研究结果表明，使用我们的方案的rpsL转基因斑马鱼突变试验是更有效的成鱼比胚胎，但移码突变可以检测到在胚胎和成人在适当的采样时间后，用ICR-191处理。(C)2005 Elsevier B. V.保留所有权利。
To determine whether frameshift mutations can be detected in rpsL transgenic zebrafish (Brachydanio rerio), embryos, and adult fish were treated with 6-chloro-9- [3-(2-chloroethylamino)-propylamino] -2-methoxyacridine (ICR-191). Embryos exposed to 0, 10, or 20 mu M ICR-191 in a water bath for 18 h exhibited induced mutant frequencies (MFs) of 14 x 10(-5), 16 x 10(-5), and 25 x 10(-5), respectively. Only embryos exposed to 20 mu M ICR- 191 showed a significant increase in ME The mutational spectra differed between the control and ICR-191-treated groups and single G:C pair insertions, which are a marked characteristic of ICR- 191 mutagenesis, were observed in both 10 and 20 mu M-treated embryos. In adult fish treated with 1 mu M ICR- 191 in a water bath for 18 h, a significant increase in MFs was observed in both gill (12 x 10-5 and 44 x 10-5 in control and treated fish, respectively), and hepatopancreas (5 x 10(-5) and 29 x 10(-5), respectively) 2 weeks after exposure. Sequence analysis showed that 58% of mutations in gill and 94% of mutations in hepatopancreas were single G:C pair insertions, which is typical of mutations induced by ICR-191. Additionally, these mutations occurred predominantly at a single site (CC sequence at bps 140-141) in the rpsL gene. Three weeks after exposure, however, the increased MFs and prominent mutational spectra of ICR-treated fish were undetectable. These findings suggest that using our protocols the rpsL transgenic zebrafish mutation assay is more effective for adult fish than for embryos, but that frameshift mutations can be detected in both embryos and adults at appropriate sampling times after treatment with ICR-191. (C) 2005 Elsevier B.V. All rights reserved.