H1foo is essential for in vitro meiotic maturation of bovine oocytes.

Zygote (Cambridge, England) Pub Date : 2015-06-01 Epub Date: 2014-03-11 DOI:10.1017/S0967199414000021
Yan Yun, Peng An, Jing Ning, Gui-Ming Zhao, Wen-Lin Yang, An-Min Lei
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引用次数: 19

Abstract

Oocyte-specific linker histone, H1foo, is localized on the oocyte chromosomes during the process of meiotic maturation, and is essential for mouse oocyte maturation. Bovine H1foo has been identified, and its expression profile throughout oocyte maturation and early embryo development has been established. However, it has not been confirmed if H1foo is indispensable during bovine oocyte maturation. Effective siRNAs against H1foo were screened in HeLa cells, and then siRNA was microinjected into bovine oocytes to down-regulate H1foo expression. H1foo overexpression was achieved via mRNA injection. Reverse transcription polymerase chain reaction (RT-PCR) results indicated that H1foo was up-regulated by 200% and down-regulated by 70%. Based on the first polar body extrusion (PB1E) rate, H1foo overexpression apparently promoted meiotic progression. The knockdown of H1foo significantly impaired bovine oocyte maturation compared with H1foo overexpression and control groups (H1foo overexpression = 88.7%, H1foo siRNA = 41.2%, control = 71.2%; P < 0.05). This decrease can be rescued by co-injection of a modified H1foo mRNA that has escaped from the siRNA target. However, the H1e (somatic linker histone) overexpression had no effect on PB1E rate when compared with the control group. Therefore we concluded that H1foo is essential for bovine oocyte maturation and its overexpression stimulates the process.

H1foo对牛卵母细胞的体外减数分裂成熟至关重要。
卵母细胞特异性连接蛋白H1foo在卵母细胞减数分裂成熟过程中定位于卵母细胞染色体上,对小鼠卵母细胞成熟至关重要。牛H1foo已被鉴定出来,并在卵母细胞成熟和早期胚胎发育过程中确定了其表达谱。然而,H1foo在牛卵母细胞成熟过程中是否不可或缺还未得到证实。在HeLa细胞中筛选抗H1foo的有效siRNA,然后将siRNA微注射到牛卵母细胞中下调H1foo的表达。通过mRNA注射实现H1foo过表达。逆转录聚合酶链反应(RT-PCR)结果显示,H1foo基因表达量上调200%,下调70%。根据第一极体挤压率(PB1E), H1foo过表达明显促进减数分裂进程。与H1foo过表达组和对照组相比,敲低H1foo显著影响牛卵母细胞成熟(H1foo过表达= 88.7%,H1foo siRNA = 41.2%,对照组= 71.2%;P < 0.05)。这种减少可以通过共同注射从siRNA靶标中逃脱的修饰的H1foo mRNA来挽救。然而,与对照组相比,H1e(体细胞连接蛋白)过表达对PB1E率没有影响。因此,我们认为H1foo对牛卵母细胞成熟至关重要,它的过表达刺激了这一过程。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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