Imai Y, Feldman B, Schier A F, Talbot W S
Department of Developmental Biology, Beckman Center, Stanford University School of Medicine, Stanford, California 94305-5329, USA.
Genetics. 2000 May;155(1):261-72. doi: 10.1093/genetics/155.1.261.
Mutations identified in zebrafish genetic screens allow the dissection of a wide array of problems in vertebrate biology. Most screens have examined mutations induced by treatment of spermatogonial (premeiotic) cells with the chemical mutagen N-ethyl-N-nitrosourea (ENU). Treatment of postmeiotic gametes with ENU induces specific-locus mutations at a higher rate than premeiotic regimens, suggesting that postmeiotic mutagenesis protocols could be useful in some screening strategies. Whereas there is extensive evidence that ENU induces point mutations in premeiotic cells, the range of mutations induced in postmeiotic zebrafish germ cells has been less thoroughly characterized. Here we report the identification and analysis of five mutations induced by postmeiotic ENU treatment. One mutation, snh(st1), is a translocation involving linkage group (LG) 11 and LG 14. The other four mutations, oep(st2), kny(st3), Df(LG 13)(st4), and cyc(st5), are deletions, ranging in size from less than 3 cM to greater than 20 cM. These results show that germ cell stage is an important determinant of the type of mutations induced. The induction of chromosomal rearrangements may account for the elevated frequency of specific-locus mutations observed after treatment of postmeiotic gametes with ENU.
在斑马鱼遗传筛选中鉴定出的突变有助于剖析脊椎动物生物学中的一系列问题。大多数筛选研究了用化学诱变剂N-乙基-N-亚硝基脲(ENU)处理精原细胞(减数分裂前)所诱导的突变。用ENU处理减数分裂后的配子诱导特定基因座突变的速率高于减数分裂前的方案,这表明减数分裂后诱变方案在某些筛选策略中可能有用。虽然有大量证据表明ENU在减数分裂前细胞中诱导点突变,但对减数分裂后斑马鱼生殖细胞中诱导的突变范围的表征却不够充分。在此,我们报告了对减数分裂后ENU处理诱导的五个突变的鉴定和分析。一个突变,snh(st1),是涉及连锁群(LG)11和LG 14的易位。其他四个突变,oep(st2)、kny(st3)、Df(LG 13)(st4)和cyc(st5),是缺失,大小范围从小于3 cM到大于20 cM。这些结果表明生殖细胞阶段是诱导突变类型的重要决定因素。染色体重排的诱导可能解释了用ENU处理减数分裂后配子后观察到的特定基因座突变频率升高的现象。