gga-miR-218在精原干细胞减数分裂中的调控作用

IF 2.6 Q2 Medicine Mechanisms of Development Pub Date : 2020-12-01 DOI:10.1016/j.mod.2020.103636

Yingjie Wang , Lei Zhang , Wenhui Zhang , Changhua Sun , Zheyu Deng , Cai Hu , Ahmed Kamel Elsayed , Xinqi Zhou , Tingting Li , Qisheng Zuo , Xinglong Wang , Bichun Li , Ya-ni Zhang

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引用次数: 1

摘要

MicroRNAs在精子形成中起着至关重要的作用，但其具体功能尚不清楚。在这里，我们通过体内/离体实验发现gga-miR-218调节鸡精子的形成。我们构建过表达/干扰载体，在鸡精原干细胞中过表达和抑制gga-miR-218，分别对减数分裂相关基因进行单倍体检测和QRT-PCR检测，发现gga-miR-218抑制减数分裂。体内注射miR-218后，精液浓度和HE(苏木精和伊红染色)显示gga-miR-218抑制减数分裂。同时，我们通过预测软件发现gga-miR-218可以靶向Stra8，通过将gga-miR-218与Stra8的3′非翻译区荧光报告载体(野生型/突变型)共转染，可以抑制Stra8的野生型荧光活性，QRT-PCR和Western blot结果显示，gga-miR-218直接靶向Stra8的3′非翻译区，从而抑制Stra8的表达水平。最后，我们认为gga-miR-218可以直接靶向srta8并抑制精子发生。

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Regulatory functions of gga-miR-218 in spermatogonial stem cells meiosis by targeting Stra8

MicroRNAs play a crucial role in sperm formation, but its specific function remains unknown. Here, we found that gga-miR-218 regulates chicken sperm formation through in/ex vivo experiments. We constructed over-expression/interference carrier to overexpress and inhibit gga-miR-218 in chicken spermatogonial stem cells, separately, the detection of haploid and QRT-PCR of meiosis related genes revealed that gga-miR-218 inhibits meiosis. After injection of miR-218 in vivo, semen concentration and HE (Hematoxylin and Eosin staining) revealed that gga-miR-218 inhibits meiosis. Meanwhile, we discovered that gga-miR-218 could target Stra8 by prediction software which can inhibit the wild-type fluorescence activity by co-transfection of gga-miR-218 with the Stra8 3′ untranslated regions fluorescent reporter vector (wild-type/mutant), QRT-PCR and Western blot showed that gga-miR-218 inhibits the expression level of Stra8 by targeting its 3′ untranslated regions directly. Finally, we suggest that gga-miR-218 could target to srta8 directly and inhibit spermatogenesis.

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来源期刊

Mechanisms of Development 生物-发育生物学

CiteScore

3.60

自引率

0.00%

发文量

审稿时长

12.4 weeks

期刊介绍： Mechanisms of Development is an international journal covering the areas of cell biology and developmental biology. In addition to publishing work at the interphase of these two disciplines, we also publish work that is purely cell biology as well as classical developmental biology. Mechanisms of Development will consider papers in any area of cell biology or developmental biology, in any model system like animals and plants, using a variety of approaches, such as cellular, biomechanical, molecular, quantitative, computational and theoretical biology. Areas of particular interest include: Cell and tissue morphogenesis Cell adhesion and migration Cell shape and polarity Biomechanics Theoretical modelling of cell and developmental biology Quantitative biology Stem cell biology Cell differentiation Cell proliferation and cell death Evo-Devo Membrane traffic Metabolic regulation Organ and organoid development Regeneration Mechanisms of Development does not publish descriptive studies of gene expression patterns and molecular screens; for submission of such studies see Gene Expression Patterns.

期刊最新文献

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