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玻璃化冷冻过程中添加抗冻糖蛋白8对牛卵母细胞发育能力的影响。

Effect of antifreeze glycoprotein 8 supplementation during vitrification on the developmental competence of bovine oocytes.

作者信息

Liang Shuang, Yuan Bao, Kwon Jeong-Woo, Ahn Mija, Cui Xiang-Shun, Bang Jeong Kyu, Kim Nam-Hyung

机构信息

Department of Animal Science, Chungbuk National University, Cheongju, Chungbuk, Republic of Korea.

Department of Animal Science, Chungbuk National University, Cheongju, Chungbuk, Republic of Korea; Department of Laboratory Animal Center, College of Animal Sciences, Jilin University, Changchun, China.

出版信息

Theriogenology. 2016 Jul 15;86(2):485-494.e1. doi: 10.1016/j.theriogenology.2016.01.032. Epub 2016 Feb 8.

Abstract

The purpose of this study was to investigate the effect of antifreeze glycoprotein 8 (AFGP8) supplementation during vitrification on the survival, fertilization, and embryonic development of bovine oocytes and the underlying molecular mechanism(s). Survival, fertilization, early embryonic development, apoptosis, DNA double-strand breaks, reactive oxygen species levels, meiotic cytoskeleton assembly, chromosome alignment, and energy status of mitochondria were measured in the present experiments. Compared with that in the nonsupplemented group; survival, monospermy, blastocyst formation rates, and blastomere counts were significantly higher in the AFGP8-supplemented animals. Oocytes of the latter group also presented fewer double-strand breaks and lower cathepsin B and caspase activities. Rates of normal spindle organization and chromosome alignment, actin filament impairment, and mitochondrial distribution were significantly higher in the AFGP8-supplemented group. In addition, intracellular reactive oxygen species levels significantly decreased in the AFGP8-supplemented groups, maintaining a higher ΔΨm than that in the nonsupplemented group. Taken together, these results indicated that supplementation with AFGP8 during vitrification has a protective effect on bovine oocytes against chilling injury.

摘要

本研究的目的是探讨玻璃化冷冻过程中添加抗冻糖蛋白8(AFGP8)对牛卵母细胞存活、受精及胚胎发育的影响及其潜在分子机制。在本实验中,对存活、受精、早期胚胎发育、细胞凋亡、DNA双链断裂、活性氧水平、减数分裂细胞骨架组装、染色体排列以及线粒体能量状态进行了测定。与未添加组相比,添加AFGP8的动物在存活、单精受精、囊胚形成率和卵裂球数量方面显著更高。后一组的卵母细胞双链断裂也更少,组织蛋白酶B和半胱天冬酶活性更低。添加AFGP8组的正常纺锤体组织和染色体排列率、肌动蛋白丝损伤及线粒体分布显著更高。此外,添加AFGP8组的细胞内活性氧水平显著降低,与未添加组相比维持更高的线粒体膜电位(ΔΨm)。综上所述,这些结果表明玻璃化冷冻过程中添加AFGP8对牛卵母细胞免受冷损伤具有保护作用。

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