表皮生长因子和胰岛素样生长因子对水牛卵母细胞胞质成熟的影响

Effects of Epidermal Growth Factor and Insulin Growth Factor-1 on the in vitro Cytoplasmic Maturation of Buffalo Oocytes

以皮层颗粒(CG)单层分布于质膜下做为卵母细胞胞质成熟的标志,利用CG荧光染色法,研究了不同浓度表皮生长因子(EGF)和胰岛素样生长因子(IGF-1)及其组合对水牛卵母细胞胞质成熟的影响。结果发现:(1)添加不同浓度的EGF(10、20、30、50 ng/mL)都可以提高胞质的成熟率,但是组间差异不显著(P>0.05);皮质颗粒的分布随着EGF浓度的升高逐步由中间分布向皮层分布转变;(2)在成熟液中添加IGF-130 ng/mL时效果较好,能显著提高卵母细胞胞质的成熟率;皮质颗粒的分布随着IGF-1浓度的升高逐步由中间分布向皮层分布转变,在添加IGF-130 ng/mL时,在皮层分布最好,随着IGF-1量的进一步增加,皮质颗粒又向中间分布转变;(3)添加20ng/mL EGF+30 ng/mL IGF-1组卵母细胞体外成熟率高于添加30 ng/mL的IGF-1组,显著高于添加20 ng/mLEGF组和对照组(P<0.05)。由此表明,EGF和IGF-1对水牛卵母细胞体外成熟有协同作用。

The cytoplasmic maturation of oocytes was evaluated according to the monolayer distribution of cortical granule （CG） beneath the plasmalemma following fluorescent staining. Effects of epidermal growth factor （EGF） and insulin growth factor-1 （IGF-1） on the cytoplasmic maturation of buffalo oocytes were examined in this study. Results showed that all EGF at the concentration tested （10, 20, 30, 50 ng/mL） could promote the maturity of buffalo oocyte cytoplasm, the distribution pattern of CG changed gradually from middle to cortex with the concentration of EGF increased. The maturity of buffalo oocyte cytoplasm was also improved significantly by adding 30 ng/mL IGF into the maturation medium （P〈0.05）, the distribution pattern of CG changed gradually from middle to cortex as the concentration of IGF-1 increased to 30 ng/mL, and then changed gradually from cortex to middle as the concentration of IGF-1 increased to either 50 ng/mL or 100 ng/mL. More oocytes matured significantly when they were cultured in the maturation medium containing 20 ng/mL EGF and 30 ng/mL IGF-1 in comparison with them cultured in either 30 ng/mL IGF-1 or 20 ng/mL EGF groups（P〈0.05）. These results indicate that EGF and IGF-1 have a cooperative effects on the in vitro maturation of buffalo oocytes.