We have recently shown that the incubation of Xenopus laevis oocytes in procaine-containing solutions induced germinal vesicle breakdown without white spot formation and, in some cases, with the appearance of spindle and chromosomes in the cytoplasm. The present study was performed to determine whether M-phase promoting factor was involved in this unusual maturation. Procaine failed to induce maturation in the presence of 6-dimethylamino purine or roscovitine, which are both known to inhibit p34cdc2 kinase. Histone H1 kinase activity was detected in procaine-treated oocytes but it was always lower than in progesterone-treated controls. A shift in p34cdc2 was observed in oocytes that had been exposed to procaine for 16 h, but it was not detected in those exposed for 24 h. Finally, cytoplasm transfer experiments demonstrated that the maturation promoting activity that occurred in oocytes incubated in procaine for 16 h could induce maturation of recipient stage VI oocytes. This transferable activity was weaker than that from progesterone-treated controls since only 30% of the recipients underwent germinal vesicle breakdown and only a few spindles were observed, which were not always correctly located. Taken together these results demonstrate that M-phase promoting factor is involved in the procaine maturing effect despite some differences compared with progesterone-treated oocytes which might explain the particular type of maturation induced by this substance. The discovery of the mechanisms by which procaine is able to activate M-phase promoting factor might now help in the understanding of some steps in progesterone-induced maturation that have still to be elucidated.

译文

我们最近表明,在含普鲁卡因的溶液中孵育非洲爪蟾卵母细胞可诱导生发囊泡破裂,而不会形成白斑,在某些情况下,细胞质中会出现纺锤体和染色体。进行本研究是为了确定M期促进因子是否参与了这种异常成熟。普鲁卡因在存在6-二甲基氨基嘌呤或roscovitine的情况下未能诱导成熟,这两者均已知会抑制p34cdc2激酶。在普鲁卡因处理的卵母细胞中检测到组蛋白H1激酶活性,但始终低于孕酮处理的对照组。在暴露于普鲁卡因16小时的卵母细胞中观察到p34cdc2的变化,但在暴露24小时的卵母细胞中未检测到p34cdc2的变化。最后,细胞质转移实验表明,在普鲁卡因中孵育16小时的卵母细胞中发生的促进成熟的活性可以诱导受体VI期卵母细胞的成熟。这种可转移活性比黄体酮治疗的对照组更弱,因为只有30% 的接受者经历了生发囊泡破裂,并且仅观察到几个纺锤体,这些纺锤体并不总是正确定位。综上所述,这些结果表明,尽管与孕酮处理的卵母细胞相比存在一些差异,但M期促进因子参与了普鲁卡因的成熟作用,这可能解释了该物质诱导的特定成熟类型。普鲁卡因能够激活M期促进因子的机制的发现现在可能有助于理解孕酮诱导的成熟中的一些步骤,这些步骤仍有待阐明。

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