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玻璃化对小鼠生发泡卵母细胞体外成熟及极光激酶A蛋白表达的影响

The impact of vitrification on murine germinal vesicle oocyte In vitro maturation and aurora kinase A protein expression.

作者信息

Doyle Joseph O, Lee Ho Joon, Selesniemi Kaisa, Styer Aaron K, Rueda Bo R

机构信息

Vincent Center for Reproductive Biology, Thier 9, MGH Vincent Department of Obstetrics and Gynecology, Massachusetts General Hospital, 55 Fruit Street, Boston, MA, 02114, USA.

出版信息

J Assist Reprod Genet. 2014 Dec;31(12):1695-702. doi: 10.1007/s10815-014-0336-7. Epub 2014 Oct 16.

Abstract

PURPOSE

Investigate the effect of vitrification on in vitro maturation (IVM) and expression of Aurora kinases A, B, and C in germinal vesicle (GV)-stage oocytes.

METHODS

GV-stage oocytes from B6D2F1 female mice 7-11 weeks of age were vitrified after collection, thawed, and matured in vitro for 0, 4, 8, and 12 h (hrs). The rate of germinal vesicle breakdown (GVBD), spindle apparatus assembly, and Aurora kinase mRNA and protein expression during IVM was measured.

RESULTS

Oocyte vitrification was associated with significant delays in both GVBD and normal spindle apparatus assembly at 4 and 8 h of IVM (p < 0.05). There was no difference in mRNA levels between control and vitrified oocytes for any of the Aurora kinases. Aurora A protein levels were reduced in vitrified compared to control oocytes at 0 h (p = 0.008), and there was no difference at 4 and 8 h (p = 0.08 and 0.69, respectively) of IVM.

CONCLUSIONS

Vitrified oocytes have delayed GVBD and normal spindle assembly during in vitro maturation. Reduced levels of Aurora A protein immediately post-thaw may be associated with the impaired oocyte maturation manifested by the delayed progression through meiosis I and II, and the atypical timing of the formation of meiotic spindles in vitrified GV-stage oocytes.

摘要

目的

研究玻璃化冷冻对生发泡(GV)期卵母细胞体外成熟(IVM)及极光激酶A、B和C表达的影响。

方法

收集7 - 11周龄B6D2F1雌性小鼠的GV期卵母细胞,收集后进行玻璃化冷冻,解冻后分别在体外成熟0、4、8和12小时(h)。检测体外成熟过程中生发泡破裂(GVBD)率、纺锤体组装情况以及极光激酶mRNA和蛋白表达。

结果

卵母细胞玻璃化冷冻与体外成熟4小时和8小时时GVBD及正常纺锤体组装的显著延迟相关(p < 0.05)。对于任何一种极光激酶,对照卵母细胞和玻璃化冷冻卵母细胞的mRNA水平均无差异。与对照卵母细胞相比,玻璃化冷冻卵母细胞在体外成熟0小时时极光A蛋白水平降低(p = 0.008),在体外成熟4小时和8小时时无差异(分别为p = 0.08和0.69)。

结论

玻璃化冷冻卵母细胞在体外成熟过程中GVBD和正常纺锤体组装延迟。解冻后即刻极光A蛋白水平降低可能与减数分裂I和II进程延迟以及玻璃化冷冻GV期卵母细胞减数分裂纺锤体形成时间异常所表现出的卵母细胞成熟受损有关。

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Effect of vitrification and thawing on human oocyte ATP concentration.玻璃化冷冻和解冻对人卵母细胞 ATP 浓度的影响。
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