Microtubule assembly and in vitro development of bovine oocytes with increased intracellular glutathione level prior to vitrification and in vitro fertilization
Microtubule assembly and in vitro development of bovine oocytes with increased intracellular glutathione level prior to vitrification and in vitro fertilization
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DOI：
10.1017/s0967199413000105
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发表时间：
2014-11-01
期刊：
影响因子：
1.7
通讯作者：
Hochi, S.
中科院分区：
文献类型：
--
作者：
Hara, H.;Yamane, I.;Hochi, S.
玻璃化冷冻是一种有效的牛卵母细胞保存技术，但其囊胚发育率仍较低。我们最近报道了一种新型的冷冻损伤，多星状体的形成，其中原核迁移和玻璃化加温和体外受精的牛卵母细胞的发育受损。本研究旨在探讨玻璃化冷冻牛卵母细胞中谷胱甘肽（GSH）含量对多星状体形成及体外发育的影响。牛卵丘卵母细胞复合体在体外成熟过程中与β-巯基乙醇（β ME）和L-半胱氨酸（Cys）的治疗导致2.5倍高的GSH含量不仅在新鲜的控制，而且在玻璃化加热的卵母细胞。在所有四个组（有或没有β ME/Cys x新鲜对照或玻璃化冷冻）中，正常受精的受精卵显示精子星的百分比>95%。玻璃化冷冻卵母细胞（比新鲜对照卵母细胞高3倍）中多个星形细胞形成的频率不受β ME/Cys细胞内GSH水平增加的影响。因此，原核的迁移和发育以及玻璃化冷冻-加温卵母细胞的囊胚产量（17对41%）没有得到改善。此外，在新鲜对照组中，GSH水平的增加对囊胚产量没有影响。
Although vitrification is a useful technique for preservation of bovine oocytes, the yield of blastocysts derived from the vitrified oocytes is still low. We have recently reported a new type of cryoinjury, multiple aster formation, by which pronuclear migration and development of vitrified-warmed and in vitro-fertilized bovine oocytes are impaired. The aim of the present study was to investigate the effect of glutathione (GSH) content of vitrified bovine oocytes on multiple aster formation and subsequent in vitro development. Treatment of bovine cumulus-oocyte complexes with beta-mercaptoethanol (beta ME) and L-cysteine (Cys) during in vitro maturation resulted in 2.5-fold higher GSH content not only in fresh control but also in vitrified-warmed oocytes. The percentage of normally fertilized zygotes exhibiting sperm aster(s) was >95% in all four groups (with or without beta ME/Cys x fresh control or vitrified). The frequency of multiple aster formation in vitrified oocytes (three-fold higher than that in fresh control oocytes) was not affected by the increased level of intracellular GSH with beta ME/Cys. Consequently, the migration and development of pronuclei as well as the yield of blastocysts from vitrified-warmed oocytes (17 versus 41%) were not improved. In addition, there was no effect of increased GSH level on the yield of blastocysts in fresh control groups.