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Expression pattern of Ngn3 in dairy goat testis and its function in promoting meiosis by upregulating Stra8

机译:Ngn3在奶山羊睾丸中的表达模式及其通过上调Stra8促进减数分裂的功能

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Objectives: Ngn3 is a typical transcription factor and marker of differentiating spermatogonial stem cells (SSCs) in mouse, belonging to the basic helix-loop-helix (bHLH) family. Its gene is specifically expressed in A type spermatogonia in mouse testis, thus plays a critical role in controlling differentiation of SSCs. However, roles of Ngn3 and its protein in dairy goat testis remain unknown. Materials and methods: Testis development and expression patterns of Ngn3 were analysed by immunofluorescence and quantitative reverse transcription-polymerase chain reaction (QRT-PCR) in the dairy goat. Furthermore, effects of its overexpression on male germline stem cells (mGSCs) were evaluated by QRT-PCR, immunofluorescence, luciferase reporter assay and western blotting. Results: Revealed that Ngn3 was expressed more highly during puberty and in the adult than in testis of other ages. Overexpression of Ngn3 promoted expression of meiosis-related gene Stra8 and stem-cell differentiation marker CD117, but suppressed expression of Plzf, a classical marker of SSCs. Furthermore, Ngn3 did not promote expression of Stra8 directly as shown in transcription and translation levels detected by luciferase reporter assay and western blotting. Conclusions: Taken together, these results suggest that Ngn3 plays an important role in spermatogenesis and that overexpression of Ngn3 can promote meiosis in testis of the dairy goat.
机译:目的:Ngn3是一种典型的转录因子,是小鼠精原干细胞(SSCs)分化的标志物,属于基本的螺旋-环-螺旋(bHLH)家族。它的基因在小鼠睾丸的A型精原细胞中特异性表达,因此在控制SSC分化中起关键作用。然而,Ngn3及其蛋白在奶山羊睾丸中的作用仍然未知。材料与方法:通过免疫荧光和定量逆转录-聚合酶链反应(QRT-PCR)分析了奶山羊睾丸的Ngn3发育和表达模式。此外,通过QRT-PCR,免疫荧光,荧光素酶报告基因分析和Western印迹评估了其过表达对雄性生殖干细胞(mGSC)的影响。结果:揭示了Ngn3在青春期和成人中的表达高于其他年龄的睾丸。 Ngn3的过表达促进了减数分裂相关基因Stra8和干细胞分化标记CD117的表达,但抑制了SSCs的经典标记Plzf的表达。此外,Ngn3不能直接促进Stra8的表达,如通过荧光素酶报告基因检测和Western印迹检测到的转录和翻译水平所示。结论:综上所述,这些结果表明Ngn3在精子发生中起重要作用,并且Ngn3的过表达可促进乳山羊睾丸的减数分裂。

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