Exogenous dibutyryl cAMP affects meiotic maturation via protein kinase A activation; it stimulates further embryonic development including blastocyst quality in pigs
Exogenous dibutyryl cAMP affects meiotic maturation via protein kinase A activation; it stimulates further embryonic development including blastocyst quality in pigs
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DOI：
10.1016/j.theriogenology.2007.09.024
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发表时间：
2008-02-01
期刊：
影响因子：
2.8
通讯作者：
Koo, D. B.
中科院分区：
文献类型：
--
作者：
Kim, J. S.;Cho, Y. S.;Koo, D. B.
生发囊泡卵母细胞中高浓度的环AMP通常会抑制GVBD。因此，维持生长中卵母细胞的 GV 阶段对于卵子的发育能力至关重要。在本研究中，我们追踪了二丁酰环 AMP 对猪减数分裂成熟和早期胚胎发育的影响。我们还研究了受 dbcAMP 影响的几种囊胚品质，包括结构完整性、线粒体膜电位和细胞凋亡。为了确定 cAMP 浓度增加是否会抑制 GVBD，我们探索了猪卵母细胞的减数分裂模式和成熟过程。当用 dbcAMP 处理 22 小时时，91.1% 的卵母细胞在 GV 阶段停滞，而对照组中只有 38.8% 的卵母细胞停滞（P < 0.05）。 IVM 完成后，dbcAMP 处理的卵母细胞处于中期 11 的比例高于未处理的卵母细胞（91.3% vs. 72.8%，P < 0.05）。蛋白质印迹分析显示，与未处理的对照相比，在 IVM 的前 22 小时内用 dbcAMP 处理的猪卵母细胞中 MPF 和 MAP 激酶活性降低（22 小时）和/或增加（44 小时）。我们还证实，dbcAMP 处理的卵母细胞中蛋白激酶 A 活性增加，表明细胞内 cAMP 浓度升高。 IVF后，dbcAMP治疗组多精受精频率较对照组降低（22.4% vs. 47.4%，P < 0.05）。此外，在 IVF 和 SCNT 胚胎中，dbcAMP 可以增强和/或减少囊胚形成、囊胚细胞数量、线粒体膜电位和细胞凋亡。我们得出的结论是，通过 dbcAMP 处理同步减数分裂恢复，通过增加线粒体膜电位并降低植入前阶段猪胚胎的细胞凋亡发生率，提高了 IVF 和 SCNT 胚胎的发育能力和胚胎质量。 (C) 2008 Elsevier Inc. 保留所有权利。
High concentrations of cyclic AMP in germinal vesicle oocytes generally inhibit GVBD. Thus, maintaining the GV stage in growing oocytes is essential for the developmental competence of the eggs. In this study, we traced the effects of dibutyryl cyclic AMP on meiotic maturation and early embryonic development in pigs. We also investigated several blastocyst qualities, including structural integrity, mitochondrial membrane potential, and apoptosis, which are affected by dbcAMP. To determine whether increased concentrations of cAMP inhibit GVBD, we explored the meiotic patterns and during maturation of pig oocytes. When treated with dbcAMP for 22 h, 91.1% of the oocytes were arrested in the GV stage compared to only 38.8% of the oocytes in the control group (P < 0.05). After completion of IVM, a higher proportion of the dbcAMP-treated oocytes were in metaphase 11 than the untreated ones (91.3% vs. 72.8%, P < 0.05). Western blot analysis showed a reduction (at 22 h) and/or increase (at 44 h) in MPF and MAP kinase activities in porcine oocytes treated with dbcAMP for the first 22 h of IVM compared to the untreated control. We also confirmed that protein kinase A activity increased in dbcAMP-treated oocytes, indicating an elevated intracellular concentration of cAMP. After IVF, the frequency of polyspermy in the dbcAMP-treated group decreased compared to that in the control group (22.4% vs. 47.4%, P < 0.05). Furthermore, blastocyst formation, the blastocyst cell number, mitochondrial membrane potential, and apoptosis were enhanced and/or reduced by dbcAMP in both IVF and SCNT embryos. We concluded that synchronizing meiotic resumption by dbcAMP treatment improved the developmental capacity and embryonic qualities of IVF and SCNT embryos by increasing the mitochondrial membrane potential and decreasing the incidence of apoptosis in preimplantation-stage porcine embryos. (C) 2008 Elsevier Inc. All rights reserved.