Xiong Xian-Rong, Lan Dao-Liang, Li Jian, Lin Ya-Qiu, Li Ming-Yang
College of Horticulture and Landscape Architecture, Southwest University, Chongqing 400716, China; College of Life Science and Technology, Southwest Minzu University, Chengdu, Sichuan 610041, China.
College of Life Science and Technology, Southwest Minzu University, Chengdu, Sichuan 610041, China.
Anim Reprod Sci. 2017 Nov;186:21-30. doi: 10.1016/j.anireprosci.2017.08.013. Epub 2017 Aug 24.
The efficiency of in vitro embryo production remains low compared with that observed in vivo. Recent studies have independently shown that cyclic adenosine monophosphate (cAMP) modulation prior to in vitro maturation (IVM) supplementation improves oocyte developmental competence. In this context, special cAMP modulators have been applied during IVM as promising alternatives to improve this biotechnology. Accordingly, this study was conducted to evaluate the effects of treatment with cilostazol, a PDE3 inhibitor, during pre-IVM culture on oocyte meiotic maturation in yak. Immature yak cumulus-oocyte complexes (COCs) were treated in vitro without (control) or with 5μM cilostazol for 0, 2, or 4h prior to IVM. Results showed that the presence of cilostazol in pre-IVM medium significantly increased the percentages of oocytes at metaphase II stage compared with that in the control groups (P<0.05). Moreover, pre-IVM with cilostazol significantly enhanced intraoocyte cAMP and glutathione (GSH) levels at the pre-IVM or IVM phase relative to the no pre-IVM groups (P<0.05). After in vitro fertilization (IVF) and parthenogenetic activation (PA), the developmental competences of oocytes and embryo quality were improved significantly after pre-IVM with cilostazol compared with the control groups (P<0.05), given that the cleavage and blastocyst formation rates and the total number of blastocyst cells were increased. The presence of cilostazol also increased the levels of mRNA expression for adenylate cyclase 3 (ADCY3) and protein kinase 1 (PKA1), as well as decreased the abundance of phosphodiesterase 3A (PDE3A) in COCs and IVF blastocysts, compared with their control counterparts (P<0.05). The results demonstrated that the meiotic progression of immature yak oocytes could be reversibly affected by cAMP modulators. By contrast, treatment with cilostazol during pre-IVM positively affected the developmental competence of yak oocytes, probably by improving intraoocyte cAMP and GSH levels and regulating mRNA expression patterns. We concluded that appropriate treatment with cilostazol during pre-IVM would be beneficial for oocyte maturation in vitro.
与体内观察到的情况相比，体外胚胎生产效率仍然较低。最近的研究独立表明，在体外成熟（IVM）补充之前进行环磷酸腺苷（cAMP）调节可提高卵母细胞的发育能力。在这种情况下，特殊的cAMP调节剂已在IVM过程中作为有前景的替代方法应用，以改进这种生物技术。因此，本研究旨在评估在牦牛IVM前培养期间用西洛他唑（一种PDE3抑制剂）处理对卵母细胞减数分裂成熟的影响。未成熟的牦牛卵丘-卵母细胞复合体（COCs）在IVM前体外分别不处理（对照组）或用5μM西洛他唑处理0、2或4小时。结果表明，与对照组相比，IVM前培养基中存在西洛他唑显著增加了处于中期II期的卵母细胞百分比（P<0.05）。此外，与未进行IVM前处理的组相比，IVM前用西洛他唑处理在IVM前或IVM阶段显著提高了卵母细胞内cAMP和谷胱甘肽（GSH）水平（P<0.05）。在体外受精（IVF）和孤雌激活（PA）后，与对照组相比，IVM前用西洛他唑处理后卵母细胞的发育能力和胚胎质量显著提高（P<0.05），因为卵裂率、囊胚形成率和囊胚细胞总数增加。与对照相比，西洛他唑的存在还增加了COCs和IVF囊胚中环化酶3（ADCY3）和蛋白激酶1（PKA1）的mRNA表达水平，并降低了磷酸二酯酶3A（PDE3A）的丰度（P<0.05）。结果表明，未成熟牦牛卵母细胞的减数分裂进程可能受到cAMP调节剂的可逆影响。相比之下，IVM前用西洛他唑处理对牦牛卵母细胞的发育能力有积极影响，可能是通过提高卵母细胞内cAMP和GSH水平以及调节mRNA表达模式。我们得出结论，IVM前适当用西洛他唑处理有利于体外卵母细胞成熟。
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