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早幼粒细胞白血病锌指蛋白维持雄性生殖干细胞(mGSCs)的自我更新及其在奶山羊睾丸中的表达模式。

Promyelocytic leukaemia zinc finger maintains self-renewal of male germline stem cells (mGSCs) and its expression pattern in dairy goat testis.

机构信息

College of Veterinary Medicine, Shaanxi Centre of Stem Cells Engineering & Technology, Key Lab for Animal Biotechnology of Agriculture Ministry of China, Northwest A&F University, Yangling, Shaanxi, 712100, China.

出版信息

Cell Prolif. 2013 Aug;46(4):457-68. doi: 10.1111/cpr.12048.

Abstract

OBJECTIVES

Previous studies have shown that promyelocytic leukaemia zinc finger (PLZF) is a spermatogonia-specific transcription factor in the testis, required to regulate self-renewal and maintenance of the spermatogonia stem cell. Up to now, expression and function of PLZF in the goat testis has not been known. The objectives of this study were to investigate PLZF expression pattern in the dairy goat and its effect on male goat germline stem cell (mGSC) self-renewal and differentiation.

MATERIALS AND METHODS

Testis development and expression patterns of PLZF in the dairy goat were analysed by haematoxylin and eosin staining, immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, effects of PLZF overexpression on mGSC self-renewal and differentiation were evaluated by quantitative RT-PCR (QRT-PCR), immunofluorescence and BrdU incorporation assay.

RESULTS

Promyelocytic leukaemia zinc finger was essential for dairy goat testis development and expression of several proliferation and pluripotency-associated proteins including OCT4, C-MYC were upregulated by PLZF overexpression. The study demonstrated that PLZF played a key role in maintaining self-renewal of mGSCs and its overexpression enhanced expression of proliferation-associated genes.

CONCLUSIONS

Promyelocytic leukaemia zinc finger could function in the dairy goat as well as in other species in maintaining self-renewal of germline stem cells and this study provides a model to study the mechanism on self-renewal and differentiation of mGSCs in livestock.

摘要

目的

先前的研究表明,早幼粒细胞白血病锌指(PLZF)是睾丸中精原细胞特异性转录因子,需要调节精原干细胞的自我更新和维持。迄今为止,PLZF 在山羊睾丸中的表达和功能尚不清楚。本研究的目的是研究 PLZF 在奶山羊中的表达模式及其对雄性山羊生殖干细胞(mGSC)自我更新和分化的影响。

材料和方法

通过苏木精和伊红染色、免疫组织化学和逆转录-聚合酶链反应(RT-PCR)分析睾丸发育和 PLZF 在奶山羊中的表达模式。此外,通过定量 RT-PCR(QRT-PCR)、免疫荧光和 BrdU 掺入测定评估 PLZF 过表达对 mGSC 自我更新和分化的影响。

结果

早幼粒细胞白血病锌指对于山羊睾丸发育和几种增殖和多能性相关蛋白(包括 OCT4、C-MYC)的表达至关重要,PLZF 过表达上调了这些蛋白的表达。研究表明,PLZF 在维持 mGSCs 的自我更新中发挥着关键作用,其过表达增强了增殖相关基因的表达。

结论

PLZF 可以在奶山羊以及其他物种中维持生殖干细胞的自我更新,本研究为研究家畜 mGSCs 自我更新和分化的机制提供了模型。

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