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Akt 在体内和体外成熟的小鼠卵母细胞中的表达。

Akt expression in mouse oocytes matured in vivo and in vitro.

机构信息

Department of Health Sciences, University of L'Aquila, Via Vetoio, 67100 L'Aquila, Italy.

出版信息

Reprod Biomed Online. 2010 Jan;20(1):35-41. doi: 10.1016/j.rbmo.2009.10.011. Epub 2009 Oct 30.

Abstract

To improve developmental competence of in vitro matured oocytes, culture medium can be supplemented with hypoxanthine (Hx) and FSH or epidermal growth factor (EGF) to trigger the activation of essential signalling pathways regulating meiotic resumption and progression. Since the serine/threonine kinase, Akt, contributes to the regulation of the meiotic cell cycle, this study analysed its expression level and localization at the meiotic spindle in oocytes matured in vivo or in vitro in the presence of Hx-FSH or Hx-EGF. Independently of culture conditions adopted, Akt mRNA concentration did not vary from germinal vesicle to metaphase I (MI), while at MII a significant decrease in Akt1 mRNA concentration was recorded in oocytes matured in vivo and in those stimulated by Hx-EGF (P < 0.05). Phoshorylated Akt protein content was similar in the different groups of MI oocytes, but it decreased at MII in oocytes matured either in vivo or in vitro with Hx-EGF. Ser-473-phosphorylated Akt was localized uniformly to the meiotic spindle in more than 90% of oocytes. These results indicate that, in mouse oocytes, Akt expression is differentially regulated during in vivo and in vitro maturation and suggest that EGF could be a positive modulator, even stronger than FSH, of oocyte meiotic maturation.

摘要

为了提高体外成熟卵母细胞的发育能力,可以在培养基中添加次黄嘌呤(Hx)和 FSH 或表皮生长因子(EGF),以触发调节减数分裂恢复和进展的基本信号通路的激活。由于丝氨酸/苏氨酸激酶 Akt 有助于调节减数分裂细胞周期,因此本研究分析了其在体内或体外成熟的卵母细胞中在 Hx-FSH 或 Hx-EGF 存在下的表达水平和在减数分裂纺锤体中的定位。无论采用何种培养条件,Akt mRNA 浓度从生发泡到中期 I(MI)都没有变化,而在 MII 期,体内成熟和 Hx-EGF 刺激的卵母细胞中 Akt1 mRNA 浓度显著降低(P < 0.05)。不同 MI 卵母细胞组的磷酸化 Akt 蛋白含量相似,但在体内或体外与 Hx-EGF 成熟的卵母细胞中,MII 期的含量下降。Ser-473 磷酸化 Akt 均匀定位于超过 90%的卵母细胞的减数分裂纺锤体中。这些结果表明,在小鼠卵母细胞中,Akt 表达在体内和体外成熟过程中受到差异调节,并表明 EGF 可能是卵母细胞减数分裂成熟的正调节剂,甚至比 FSH 更强。

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