Isolation and cytogenetic characterization of zebrafish meiotic prophase I mutants
Isolation and cytogenetic characterization of zebrafish meiotic prophase I mutants
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DOI：
10.1002/dvdy.22661
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发表时间：
2011-07
影响因子：
2.5
通讯作者：
Kenji Saito;K. Siegfried;C. Nüsslein-Volhard;Noriyoshi Sakai
中科院分区：
文献类型：
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作者：
Kenji Saito;K. Siegfried;C. Nüsslein-Volhard;Noriyoshi Sakai
在这里，我们描述了三个减数分裂前期I突变体的分离和细胞遗传学特征，分别命名为ietsugu(ITS)，iesada(Isa)和iemochi(Imo)，这三个突变体是通过一个新的N-乙基-N-亚硝基斑马鱼性腺发生突变筛选而分离出来的。对这些突变体的组织学检查和流式细胞仪分析表明，每个突变体都没有精子细胞，也没有精子。Sycp3和裂解Caspase-3的染色和TUNEL(末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记)进一步显示其在减数分裂开始时存在缺陷，ISA和IMO精母细胞未能通过合子期，睾丸体细胞发生凋亡。组蛋白H_(2AX)磷酸化染色显示，ISA和IMO组细线精母细胞内的病灶形成被破坏。此外，体外分化实验表明，与突变相关的缺陷和不育可能是生殖系自主的。因此，我们的结果表明，在精子发生和雄性生育过程中，每个负责基因都是减数分裂进程所必需的。《发展动力学240：1779-1792》，2011年。©2011 Wiley-Liss，Inc.
We describe here the isolation and cytogenetic characterization of three meiotic prophase I mutants, denoted ietsugu (its), iesada (isa), and iemochi (imo), isolated by a novel N‐ethyl‐N‐nitrosourea mutagenesis screen for adult zebrafish gonadogenesis. Histological examination and flow cytometry analysis of testes from these mutants showed that each contained neither spermatids nor sperm. Staining for Sycp3 and cleaved Caspase‐3 and TUNEL (terminal deoxynucleotidyl transferase–mediated deoxyuridinetriphosphate nick end‐labeling) assay further revealed that its had defects at the onset of meiosis, and that isa and imo spermatocytes failed to progress past the zygotene stage with apoptosis occurring in the testicular somatic cells. Staining for phosphorylated histone H2AX showed that foci formation in leptotene spermatocytes was disrupted in isa and imo. Furthermore, in vitro differentiation experiments revealed the possibility that the defects and sterility associated with mutations were germ line autonomous. Our results thus indicate that each responsible gene is necessary for meiotic progression during spermatogenesis and for male fertility. Developmental Dynamics 240:1779–1792, 2011. © 2011 Wiley‐Liss, Inc.