蛋白激酶Pkmyt1对小鼠1-细胞期受精卵发育的抑制作用

doi:10.13481/j.1671-587x.20150104

Abstract

Abstract:

Objective To study the influence of protein kinase Pkmyt1 on the development of one-cell stage mouse embryos by the means of overexpression of Pkmyt1-wild type (WT) mRNAs, and to pay foundation for further study on the mechanism of the early development of mouse embryos. Methods pcDNA3.1/myc-Pkmyt1-WT was constructed and one-cell-stage mouse embryos were collected after superovulation, and the embryos were cultured in vitro and divided into control group(no injection), TE buffer microinjection group and Pkmyt1-WT mRNAs-injected group.The protein expression levels of Pkmyt1 and the phosphorylation status of Cdc2-pTyr15 were observed by Western blotting method.The morphological changes and cleavage rates of mouse embryos were observed under phase-contrast microscope. Results pcDNA3.1/myc-Pkmyt1-WT was constructed successfully.The Pkmyt1 protein expression level was increased at 4 h after Pkmyt1-WT-mRNAs microinjection into one-cell stage mouse embryos (1.112±0.025)compared with no injection group(0.414±0.016) and TE buffer injection group(0.441±0.013)(P<0.01).The cleavage rate of one-cell-stage mouse embryos in Pkmyt1-WT mRNAs group (34.2%±3.25%) was significantly lower than those in no injection group(60.81%±3.27%) and TE buffer injection group(61.79%±4.08%)(P<0.01).The cdc2-pTyr15 phosphorylation signal in Pkmyt1-WT-mRNA-injected group was detected at 29.0 h after hCG injection, and there was no signal at 29.5 h. Conclusion Overexpression of Pkmyt1 phosphorylates Cdc2-pTyr15, thus inhibits the cleavage rate of one-cell-stage mouse embryos, suggesting that Pkmyt1 negatively regulates mitosis during early embryogenesis in mouse embryos.

Key words: protein kinase Pkmyt1, protein kinase Wee1, mitosis, mouse embryos

CLC Number:

Q132

LIU Chao, MENG Zhichao, REN Lili, LIU Yimeng, GUO Miao, SUN Yiya, XIAO Jianying. Inhibitory effect of protein kinase Pkmyt1 on development of one-cell-stage mouse embryos[J].Journal of Jilin University Medicine Edition, 2015, 41(01): 16-20.

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Inhibitory effect of protein kinase Pkmyt1 on development of one-cell-stage mouse embryos

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