Tan Ya-Jing, Zhang Xue-Ying, Ding Guo-Lian, Li Rong, Wang Li, Jin Li, Lin Xian-Hua, Gao Ling, Sheng Jian-Zhong, Huang He-Feng
Center of Reproductive Medicine, the International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Institute of Embryo-Fetal Original Adult Disease Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai Jiao Tong University, Shanghai, China.
Sci Rep. 2015 Dec 4;5:17741. doi: 10.1038/srep17741.
Hyperosmotic stress may induce apoptosis of different cells. However, oocytes show tolerance to osmotic stress during cryopreservation by vitrification, which is an assisted reproductive technique. The underlying mechanism is still not understood. Here, we demonstrated that hyperosmosis produced by high concentrations of cryoprotectants, including DMSO, ethylene glycol and sucrose, significantly upregulated the protein levels of aquaporin (AQP) 7, but not AQP3 and AQP9, in mouse oocytes. Knockdown of AQP7 expression by siRNA-injection significantly reduced the survival of oocytes after vitrification. In oocytes, AQP7 was shown to bind with F-actin, a protein involved in almost all biological events. Moreover, we found that hyperosmosis could upregulate the phosphorylation levels of CPE-binding protein (CPEB) and Aurora A. Inhibition of the PI3K and PKC pathways blocked the hyperosmosis-induced upregulation of AQP7 and the phosphorylation of CPEB and Aurora A in oocytes. In conclusion, hyperosmosis could upregulate the expression of AQP7 via Aurora A/CPEB phosphorylation mediated by the PI3K and PKC pathways, and upregulation of AQP7 plays an important role in improving of tolerance to hyperosmotic stress and survival of oocytes during cryopreservation by vitrification.
高渗应激可能诱导不同细胞发生凋亡。然而,卵母细胞在玻璃化冷冻保存(一种辅助生殖技术)过程中对渗透应激表现出耐受性。其潜在机制仍不清楚。在此,我们证明,包括二甲亚砜、乙二醇和蔗糖在内的高浓度冷冻保护剂所产生的高渗作用,显著上调了小鼠卵母细胞中水通道蛋白(AQP)7的蛋白水平,但未上调AQP3和AQP9的蛋白水平。通过注射小干扰RNA敲低AQP7的表达,显著降低了玻璃化冷冻后卵母细胞的存活率。在卵母细胞中,AQP7被证明与F-肌动蛋白结合,F-肌动蛋白几乎参与所有生物学事件。此外,我们发现高渗作用可上调CPE结合蛋白(CPEB)和Aurora A的磷酸化水平。抑制PI3K和PKC信号通路可阻断高渗作用诱导的卵母细胞中AQP7上调以及CPEB和Aurora A的磷酸化。总之,高渗作用可通过PI3K和PKC信号通路介导的Aurora A/CPEB磷酸化上调AQP7的表达,而AQP7的上调在提高卵母细胞对高渗应激的耐受性及玻璃化冷冻保存期间的存活率方面发挥重要作用。