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Maternal Cytokines CXCL12, VEGFA, and WNT5A Promote Porcine Oocyte Maturation via MAPK Activation and Canonical WNT Inhibition.
Frontiers in Cell and Developmental Biology ( IF 5.5 ) Pub Date : 2020-06-15 , DOI: 10.3389/fcell.2020.00578
Xin Liu 1, 2 , Yuchen Hao 1, 2 , Zhekun Li 1, 2 , Jilong Zhou 1, 2 , Hongmei Zhu 3 , Guowei Bu 1, 2 , Zhiting Liu 1, 2 , Xudong Hou 4 , Xia Zhang 3 , Yi-Liang Miao 1, 2, 5
Affiliation  

Maternal regulatory factors endow the oocyte with developmental competence in vivo, which might be absent in current in vitro maturation (IVM) systems, thereby compromising oocyte quality. In the present study, by employing RNA sequencing data analysis, we expect to identify potential contributing factors to support porcine oocyte maturation through binding to their receptors on the oolemma. Here, C-X-C motif chemokine ligand 12 (CXCL12), vascular endothelial growth factor A (VEGFA), and Wingless-type MMTV integration site family member 5A (WNT5A), termed CVW, are selected and confirmed to be important maternal cytokines for porcine oocyte maturation. Combined supplementation of CVW promotes the nuclear maturation percentage from 57.2% in controls to 75.9%. More importantly, these maternal cytokines improve the developmental potential of matured oocytes by parthenogenesis, fertilization, and cloning, as their blastocyst formation efficiencies and total cell numbers are increased. CVW supplementation also enlarges perivitelline space and promotes cumulus expansion, which results in a more complete transzonal projection retraction on the zona pellucida, and a reduced incidence of polyspermy in fertilized oocytes. Meanwhile, inhibiting the CVW receptor-mediated signaling pathways severely impairs oocyte meiotic resumption and cumulus expansion during IVM. We further determine that maturation improvement by CVW is achieved through activating the MAPK pathway in advance and inhibiting the canonical WNT pathway at the end of the IVM period. These findings provide a new combination of three cytokines to promote the porcine IVM process, which also holds potential to be used in human assisted reproduction technologies as well as in other species.



中文翻译:

母体细胞因子CXCL12,VEGFA和WNT5A通过MAPK激活和经典WNT抑制作用促进猪卵母细胞成熟。

孕产调节因子赋予卵母细胞发育能力 体内,目前可能不存在 体外成熟(IVM)系统,从而损害卵母细胞质量。在本研究中,我们希望通过采用RNA测序数据分析,来确定潜在的促成因素,通过与卵母细胞上的受体结合来支持猪卵母细胞的成熟。在这里,CXC主题趋化因子配体12(CXCL12),血管内皮生长因子A(VEGFA)和Wingless型MMTV整合位点家族成员5A(WNT5A)被选为CVW,并被证实是猪卵母细胞成熟的重要母体细胞因子。 。CVW的组合补充可将核成熟率从对照组的57.2%提高到75.9%。更重要的是,这些母体细胞因子通过孤雌生殖,受精和克隆提高了成熟卵母细胞的发育潜力,因为它们的胚泡形成效率和总细胞数都增加了。CVW的补充还扩大了卵周膜的空间并促进了卵丘的扩张,从而导致透明带上更完整的跨区投射回缩,并降低了受精卵母细胞中多精子的发生率。同时,抑制CVW受体介导的信号通路严重损害了IVM期间卵母细胞的减数分裂恢复和卵丘扩展。我们进一步确定,通过提前激活MAPK途径并在IVM周期结束时抑制经典WNT途径,可以实现CVW的成熟改善。这些发现提供了三种细胞因子的新组合,以促进猪IVM过程,这也具有在人类辅助生殖技术以及其他物种中使用的潜力。

更新日期:2020-07-07
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