Yao X, Tang F, Yu M, Zhu H, Chu Z, Li M, Liu W, Hua J, Peng S
College of Veterinary Medicine, Northwest Agriculture and Forestry University, Yangling, 712100, China; Shaanxi Stem Cell Engineering and Technology Research Center, Northwest Agriculture and Forestry University, Yangling, 712100, China.
Cell Prolif. 2014 Oct;47(5):396-405. doi: 10.1111/cpr.12128.
Nanos2, an RNA-binding protein, belongs to the Nanos gene-coding family and contains two CCHC zinc-finger motifs. In mouse, it plays a pivotal role in male germ cell development, and self-renewal of spermatogonial stem cells. However, little is known of its expression pattern and functions in dairy goat testis.
Immunohistochemistry and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were used to generate the expression profile of Nanos2 in dairy goat testis. Furthermore, its overexpression effects on male germline stem cells (mGSCs) were studied using qRT-PCR, immunofluorescence, dual-luciferase reporter assay and western blotting.
Nanos2 is a conservative gene expressed widely in various tissues, especially in pancreas, and it displays higher expression in adult testes than in other age groups. Overexpression of Nanos2 significantly downregulated meiosis-related genes, including Stra8 and Scp3, which induced inhibition of meiosis. Results from dual-luciferase reporter assay and western blotting indicated that Nanos2 directly downregulated Stra8 in goat GmGSCs.
Taken together, these results suggest that Nanos2 plays an important role in spermatogonia and that its overexpression restrained meiosis in the dairy goat.
Nanos2是一种RNA结合蛋白,属于Nanos基因编码家族,含有两个CCHC锌指基序。在小鼠中,它在雄性生殖细胞发育以及精原干细胞的自我更新中起关键作用。然而,其在奶山羊睾丸中的表达模式和功能却鲜为人知。
采用免疫组织化学和定量逆转录聚合酶链反应(qRT-PCR)来生成Nanos2在奶山羊睾丸中的表达谱。此外,利用qRT-PCR、免疫荧光、双荧光素酶报告基因检测和蛋白质免疫印迹法研究了其对雄性生殖系干细胞(mGSCs)的过表达效应。
Nanos2是一个保守基因,在各种组织中广泛表达,尤其是在胰腺中,并且它在成年睾丸中的表达高于其他年龄组。Nanos2的过表达显著下调了减数分裂相关基因,包括Stra8和Scp3,从而诱导减数分裂受到抑制。双荧光素酶报告基因检测和蛋白质免疫印迹法的结果表明,Nanos2在山羊GmGSCs中直接下调Stra8。
综上所述,这些结果表明Nanos2在精原细胞中起重要作用,并且其过表达抑制了奶山羊的减数分裂。