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OPS 玻璃化冷冻后未成熟与成熟牦牛卵母细胞细胞骨架基因表达与存活率比较

Cytoskeleton Genes Expression and Survival Rate Comparison Between Immature and Mature Yak Oocyte After OPS Vitrification.

机构信息

a College of Life Science and Technology , Southwest University for Nationalities , Chengdu , China.

b College of Animal Sciences and Technology , Shihezi University , Shihezi , Xinjiang , China.

出版信息

Anim Biotechnol. 2018;29(4):247-251. doi: 10.1080/10495398.2017.1369429. Epub 2017 Oct 16.

Abstract

This study was conducted to investigate the effect of vitrification on survival rate and cytoskeleton gene expression during yak oocyte maturation. The yak oocytes were incubated for 0 h [germinal vesicle (GV) stage] and in vitro matured for 24 h [metaphase II (MII) stage] to obtain immature and mature oocytes. Survival rate after vitrification were compared between immature and mature yak oocytes and cytoskeleton-related genes [cytokeratin 8 (CK8), β-actin (ACTB), and gap junction protein, alpha 1 (GJA1)] were tested by real-time PCR. Our results showed that MII stage survival rate after open pulled straw vitrification (35.60%) is significantly higher than GV stage (25.90%) oocytes. Furthermore, expression of CK8, ACTB, and GJA1 in MII stage oocytes are also significantly higher than GV stage oocytes. In conclusion, our study demonstrated that higher expression of GJA1, CK8, and ACTB in vitrify-warmed MII stage oocytes when compared with GV stage oocytes and such discrepancy might result in higher survival rate in vitrify-warmed MII stage oocytes.

摘要

本研究旨在探讨玻璃化对牦牛卵母细胞成熟过程中存活率和细胞骨架基因表达的影响。将牦牛卵母细胞分别培养 0 小时(生发泡期,GV 期)和体外成熟 24 小时(中期 II 期,MII 期),以获得不成熟和成熟的卵母细胞。比较了不成熟和成熟牦牛卵母细胞玻璃化后的存活率,并通过实时 PCR 检测了细胞骨架相关基因(角蛋白 8(CK8)、β-肌动蛋白(ACTB)和间隙连接蛋白α 1(GJA1))的表达。结果表明,开管式玻璃化拉 straw 法(Open pulled straw vitrification,OPS)处理后 MII 期卵母细胞的存活率(35.60%)显著高于 GV 期(25.90%)卵母细胞。此外,MII 期卵母细胞中 CK8、ACTB 和 GJA1 的表达也显著高于 GV 期卵母细胞。综上所述,本研究表明,与 GV 期卵母细胞相比,玻璃化后 MII 期卵母细胞中 GJA1、CK8 和 ACTB 的表达水平更高,这种差异可能导致玻璃化后 MII 期卵母细胞存活率更高。

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