The identification of genes with unique and essential functions in the development of the zebrafish, Danio rerio.
The identification of genes with unique and essential functions in the development of the zebrafish, Danio rerio.
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DOI：
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发表时间：
1996-12
期刊：
影响因子：
4.6
通讯作者：
P. Haffter;M. Granato;M. Brand;M. Mullins;M. Hammerschmidt;D. Kane;J. Odenthal;F. V. Eeden;Yun-Jin Jiang;C. Heisenberg;R. Kelsh;M. Furutani-Seiki;E. Vogelsang;D. Beuchle;Ursula Schach;Cosima Fabian;C. Nüsslein-Volhard
中科院分区：
文献类型：
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作者：
P. Haffter;M. Granato;M. Brand;M. Mullins;M. Hammerschmidt;D. Kane;J. Odenthal;F. V. Eeden;Yun-Jin Jiang;C. Heisenberg;R. Kelsh;M. Furutani-Seiki;E. Vogelsang;D. Beuchle;Ursula Schach;Cosima Fabian;C. Nüsslein-Volhard
在大规模筛选中，我们分离出斑马鱼胚胎或早期幼虫中表现出特定可见表型的突变体。雄性用乙基亚硝基脲（ENU）进行诱变，并饲养 F2 家族的兄弟姐妹 F1 鱼之间的单对交配，诱变基因组的杂合子。 F2 兄弟姐妹之间的几次杂交获得了产卵，从而对 3857 个突变基因组进行了评分。使用立体显微镜对 F3 后代在发育的第二天、第三天和第六天进行评分。在随后的筛选中，分析了固定胚胎的正确视网膜顶盖投影。总共鉴定出 4264 个突变体。三分之二表现出普遍异常的突变体最终被丢弃。我们保留并表征了 1163 个突变体。在具有相似表型的突变体之间进行的互补杂交中，894 个突变体已被分配给 372 个基因。平均等位基因频率为 2.4。我们鉴定了参与早期发育、脊索、大脑、脊髓、体节、肌肉、心脏、循环、血液、皮肤、鳍、眼睛、耳囊、下颌和鳃弓、色素模式、色素形成、肠道、肝脏、运动和触觉反应的基因。我们的收集包含几乎所有先前描述的斑马鱼突变体的等位基因。根据等位基因频率和其他考虑因素，我们估计突变体定义的 372 个基因可能占使用我们的筛选标准可能发现的所有基因的一半以上。在这里，我们概述了所获得的突变表型谱，并讨论了斑马鱼遗传饱和筛选的局限性和潜力。
In a large-scale screen, we isolated mutants displaying a specific visible phenotype in embryos or early larvae of the zebrafish, Danio rerio. Males were mutagenized with ethylnitrosourea (ENU) and F2 families of single pair matings between sibling F1 fish, heterozygous for a mutagenized genome, were raised. Egg lays were obtained from several crosses between F2 siblings, resulting in scoring of 3857 mutagenized genomes. F3 progeny were scored at the second, third and sixth day of development, using a stereomicroscope. In a subsequent screen, fixed embryos were analyzed for correct retinotectal projection. A total of 4264 mutants were identified. Two thirds of the mutants displaying rather general abnormalities were eventually discarded. We kept and characterized 1163 mutants. In complementation crosses performed between mutants with similar phenotypes, 894 mutants have been assigned to 372 genes. The average allele frequency is 2.4. We identified genes involved in early development, notochord, brain, spinal cord, somites, muscles, heart, circulation, blood, skin, fin, eye, otic vesicle, jaw and branchial arches, pigment pattern, pigment formation, gut, liver, motility and touch response. Our collection contains alleles of almost all previously described zebrafish mutants. From the allele frequencies and other considerations we estimate that the 372 genes defined by the mutants probably represent more than half of all genes that could have been discovered using the criteria of our screen. Here we give an overview of the spectrum of mutant phenotypes obtained, and discuss the limits and the potentials of a genetic saturation screen in the zebrafish.