Effects of Growth Differentiation Factor 9 and Bone Morphogenetic Protein 15 on the in vitro Maturation of Porcine Oocytes
Effects of Growth Differentiation Factor 9 and Bone Morphogenetic Protein 15 on the in vitro Maturation of Porcine Oocytes
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DOI：
10.1111/rda.12254
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发表时间：
2014-04-01
影响因子：
1.7
通讯作者：
Kim, N-H
中科院分区：
文献类型：
--
作者：
Lin, Z-L;Li, Y-H;Kim, N-H
内容生长分化因子9（GDF 9）和骨形态发生蛋白15（BMP 15）是转化生长因子-（TGF-）家族的成员，它们在小鼠和人类卵母细胞成熟和卵丘扩张中的作用是众所周知的，但在猪中还没有。我们研究了猪卵母细胞体外成熟过程中GDF 9和BMP 15的表达。GDF 9和BMP 15的mRNA水平在胚泡破裂时观察到显著增加，表达水平在中期I（MI）达到峰值，但在中期II（MII）降低。GDF 9和BMP 15蛋白定位于卵母细胞胞质。虽然GDF 9和BMP 15处理增加了参与卵母细胞成熟（c-mos，cyclinb 1和cdc 2）和卵丘扩张（has 2，ptgs 2，ptx 3和tnfaip 6）的基因表达，但SB 431542（TGF-GDF 9抑制剂）降低了MII的减数分裂成熟。孤雌激活后，SB 431542处理组的囊胚率低于对照组（分别为41.3%和74.4%）。GDF 9和BMP 15处理也增加了母体基因的mRNA水平，如c-mos [丝裂原活化蛋白激酶（MAPK）的调节亚基]和cyclinb 1和cdc 2 [成熟/M期促进因子（MPF）的调节亚基];然而，SB 431542显著降低了它们的mRNA水平。这些数据得到poly（A）-测试PCR和蛋白质活性分析的支持。我们的研究结果表明，GDF 9和BMP 15参与卵丘扩张，他们刺激MPF和MAPK活性在猪卵母细胞体外成熟过程中。
Contents Growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) are members of the transforming growth factor- (TGF-) family, and their roles in oocyte maturation and cumulus expansion are well known in the mouse and human, but not in the pig. We investigated GDF9 and BMP15 expressions in porcine oocytes during in vitro maturation. A significant increase in the mRNA levels of GDF9 and BMP15 was observed at germinal vesicle breakdown, with expression levels peaking at metaphase I (MI), but decreasing at metaphase II (MII). GDF9 and BMP15 protein localized to the oocyte cytoplasm. While treatment with GDF9 and BMP15 increased the expression of genes involved in both oocyte maturation (c-mos, cyclinb1 and cdc2) and cumulus expansion (has2, ptgs2, ptx3 and tnfaip6), SB431542 (a TGF-GDF9 inhibitor) decreased meiotic maturation at MII. Following parthenogenetic activation, the percentage of blastocysts in SB431542 treatment was lower than in the control (41.3% and 74.4%, respectively). Treatment with GDF9 and BMP15 also increased the mRNA levels of maternal genes such as c-mos [a regulatory subunit of mitogen-activated protein kinase (MAPK)], and cyclinb1 and cdc2 [regulatory subunits of maturation/M-phase-promoting factor (MPF)]; however, SB431542 significantly decreased their mRNA levels. These data were supported by poly (A)-test PCR and protein activity analyses. Our results show that GDF9 and BMP15 participate in cumulus expansion and that they stimulate MPF and MAPK activities in porcine oocytes during in vitro maturation.