Maternal Cytokines CXCL12, VEGFA, and WNT5A Promote Porcine Oocyte Maturation via MAPK Activation and Canonical WNT Inhibition
Maternal Cytokines CXCL12, VEGFA, and WNT5A Promote Porcine Oocyte Maturation via MAPK Activation and Canonical WNT Inhibition
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母体细胞因子 CXCL12、VEGFA 和 WNT5A 通过 MAPK 激活和典型 WNT 抑制促进猪卵母细胞成熟
DOI：
10.3389/fcell.2020.00578
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发表时间：
2020-07-07
影响因子：
5.5
通讯作者：
Miao, Yi-Liang
中科院分区：
文献类型：
--
作者：
Liu, Xin;Hao, Yuchen;Miao, Yi-Liang
母体调节因素赋予卵母细胞体内发育能力，而这在当前的体外成熟（IVM）系统中可能不存在，从而损害了卵母细胞的质量。在本研究中，通过采用RNA测序数据分析，我们期望通过与卵膜上的受体结合来识别支持猪卵母细胞成熟的潜在影响因素。在这里，C-X-C 基序趋化因子配体 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)（称为 CVW）被选择并确认为猪卵母细胞成熟的重要母体细胞因子。联合补充 CVW 可使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单性生殖、受精和克隆提高成熟卵母细胞的发育潜力，因为它们的囊胚形成效率和细胞总数增加。补充 CVW 还可以扩大卵周空间并促进卵丘扩张，从而导致透明带上更完整的跨区投射收缩，并减少受精卵母细胞中多精卵的发生率。同时，抑制 CVW 受体介导的信号通路会严重损害 IVM 期间卵母细胞减数分裂的恢复和卵丘扩张。我们进一步确定，CVW 的成熟改善是通过提前激活 MAPK 途径并在 IVM 期结束时抑制典型的 WNT 途径来实现的。这些发现提供了三种细胞因子的新组合来促进猪 IVM 过程，这也有可能用于人类辅助生殖技术以及其他物种。
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.