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序列相似性家族181成员A()基因敲除不会损害小鼠的精子发生或雄性生育能力。

Knockout of the family with sequence similarity 181, member A () gene does not impair spermatogenesis or male fertility in the mouse.

作者信息

Shah Wasim, Khan Ranjha, Shah Basit, Dil Sobia, Shi Qinghua

机构信息

The First Affiliated Hospital of the University of Science and Technology of China, Hefei National Laboratory for Physical Sciences at Microscale, School of Life Sciences, University of Science and Technology of China, Hefei 230027, China.

The First Affiliated Hospital of the University of Science and Technology of China, Hefei National Laboratory for Physical Sciences at Microscale, School of Life Sciences, University of Science and Technology of China, Hefei 230027, China; and Corresponding authors.

出版信息

Reprod Fertil Dev. 2021 Jul;33:674-681. doi: 10.1071/RD21150.

Abstract

Family with sequence similarity 181 (Fam181 ) is a gene family with two paralogues (Fam181a and Fam181b ) found among vertebrates. Fam181a exhibits dynamic and stage-specific expression during murine embryo development. Furthermore, searching in the National Center for Biotechnology Information database revealed predominant expression of Fam181a in mouse and human testes, implying that it may have essential roles in spermatogenesis. In this study we investigated the invivo function of Fam181a in mouse spermatogenesis and fertility by generating Fam181a -/- mice using clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) 9 genome editing technology. The resulting Fam181a -/- mice exhibited normal growth and development. In addition, the mice were completely fertile, with no obvious differences in the testis-to-bodyweight ratio, epididymal sperm count or sperm motility compared with wild-type mice. Further examination of testicular and epididymal histology of Fam181a -/- mice found an intact seminiferous tubule structure and the presence of all types of germ cells, from spermatogonia to mature spermatozoa, similar to wild-type littermates. Similarly, analysis of meiotic prophase I progression revealed normal populations of each substage of prophase I in Fam181a +/+ and Fam181a -/- testes, suggesting that this gene is dispensable for male fertility. These negative findings will help avoid research overlap, save time and resources and allow researchers to concentrate on genes that are critical for male fertility and spermatogenesis.

摘要

序列相似性家族181(Fam181)是一个基因家族,在脊椎动物中发现有两个旁系同源基因(Fam181a和Fam181b)。Fam181a在小鼠胚胎发育过程中表现出动态的、阶段特异性的表达。此外,在国家生物技术信息中心数据库中搜索发现,Fam181a在小鼠和人类睾丸中主要表达,这意味着它可能在精子发生中具有重要作用。在本研究中,我们使用成簇规律间隔短回文重复序列(CRISPR)/CRISPR相关蛋白9(Cas)9基因组编辑技术构建Fam181a基因敲除小鼠,以研究Fam181a在小鼠精子发生和生育能力中的体内功能。所得到的Fam181a基因敲除小鼠生长发育正常。此外,这些小鼠完全可育,与野生型小鼠相比,其睾丸与体重之比、附睾精子计数或精子活力均无明显差异。对Fam181a基因敲除小鼠的睾丸和附睾组织学进行进一步检查发现,其生精小管结构完整,存在从精原细胞到成熟精子的所有类型生殖细胞,与野生型同窝小鼠相似。同样,对减数分裂前期I进程的分析表明,Fam181a+/+和Fam181a-/-睾丸中前期I各亚阶段的细胞群体正常,这表明该基因对雄性生育能力并非必需。这些阴性结果将有助于避免研究重叠,节省时间和资源,并使研究人员能够专注于对雄性生育能力和精子发生至关重要的基因。

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