Heterozygous mutation of eEF1A1b resulted in spermatogenesis arrest and infertility in male tilapia, Oreochromis niloticus.
Heterozygous mutation of eEF1A1b resulted in spermatogenesis arrest and infertility in male tilapia, Oreochromis niloticus.
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eEF1A1b 杂合突变导致雄性罗非鱼 Oreochromis niloticus 精子发生停滞和不育
DOI：
10.1038/srep43733
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发表时间：
2017-03-07
影响因子：
4.6
通讯作者：
Wang D
中科院分区：
文献类型：
--
作者：
Chen J;Jiang D;Tan D;Fan Z;Wei Y;Li M;Wang D
真核细胞延伸因子1 α（eEF 1A）是翻译装置的重要组成部分。在本研究中，eEF 1A 1b分离自尼罗罗非鱼。Real-time PCR和Western blot结果表明，eEF 1A 1b在90 dah（孵化后天数）以后的精巢中表达量较高。原位杂交和免疫组织化学分析表明，eEF 1A 1b在睾丸精原细胞中高度表达。CRISPR/Cas9介导的eEF 1A 1b突变导致FO XY鱼的精子发生停滞和不育。因此，eEF 1A 1b杂合突变（eEF 1A 1b +/−）导致90 dah时无精母细胞，180 dah时精母细胞、精子细胞和精子数量极少，Cyp 11b 2和血清11-酮睾酮水平在两个阶段均降低。对精子受精能力的进一步检查表明，eEF 1A 1b +/-XY鱼由于精子发生异常而不育。来自180 dah XY鱼的eEF 1A 1b +/−睾丸的转录组学分析显示，与对照XY相比，涉及精子发生、类固醇生成和精子运动的关键元件显著下调。eEF 1A 1b的转基因过表达挽救了eEF 1A 1b +/−睾丸的精子发生停滞表型。综上所述，我们的数据表明，eEF 1A 1b是至关重要的精子发生和雄性生育力的尼罗罗非鱼。
Eukaryotic elongation factor 1 alpha (eEF1A) is an essential component of the translational apparatus. In the present study, eEF1A1b was isolated from the Nile tilapia. Real-time PCR and Western blot revealed that eEF1A1b was expressed highly in the testis from 90 dah (days after hatching) onwards. In situ hybridization and immunohistochemistry analyses showed that eEF1A1b was highly expressed in the spermatogonia of the testis. CRISPR/Cas9 mediated mutation of eEF1A1b resulted in spermatogenesis arrest and infertility in the F0 XY fish. Consistently, heterozygous mutation of eEF1A1b (eEF1A1b+/−) resulted in an absence of spermatocytes at 90 dah, very few spermatocytes, spermatids and spermatozoa at 180 dah, and decreased Cyp11b2 and serum 11-ketotestosterone level at both stages. Further examination of the fertilization capacity of the sperm indicated that the eEF1A1b+/− XY fish were infertile due to abnormal spermiogenesis. Transcriptomic analyses of the eEF1A1b+/− testis from 180 dah XY fish revealed that key elements involved in spermatogenesis, steroidogenesis and sperm motility were significantly down-regulated compared with the control XY. Transgenic overexpression of eEF1A1b rescued the spermatogenesis arrest phenotype of the eEF1A1b+/− testis. Taken together, our data suggested that eEF1A1b is crucial for spermatogenesis and male fertility in the Nile tilapia.