The effect of M-phase stage-dependent kinase inhibitors on inositol 1,4,5-trisphosphate receptor 1 (IP3 R1) expression and localization in pig oocytes.
The effect of M-phase stage-dependent kinase inhibitors on inositol 1,4,5-trisphosphate receptor 1 (IP3 R1) expression and localization in pig oocytes.
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DOI：
10.1111/asj.12258
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发表时间：
2015-02
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影响因子：
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通讯作者：
A. Sathanawongs;Katsuyoshi Fujiwara;Tsubasa Kato;M. Hirose;M. Kamoshita;R. Wojcikiewicz;J. Parys;J. Ito;N. Kashiwazaki
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文献类型：
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作者：
A. Sathanawongs;Katsuyoshi Fujiwara;Tsubasa Kato;M. Hirose;M. Kamoshita;R. Wojcikiewicz;J. Parys;J. Ito;N. Kashiwazaki
在受精过程中，1型肌醇1，4，5-三磷酸受体（IP 3 R1）在哺乳动物的Ca（2+）释放中起着至关重要的作用。IP 3 R1的表达水平、定位和磷酸化对其功能至关重要，但目前尚不清楚哪些分子调控IP 3 R1在猪卵母细胞中的行为。我们研究了是否有一个在体外或体内成熟的猪卵母细胞后，在本地化的IP 3 R1的差异。在小鼠卵母细胞中，大集群的IP 3 R1形成在卵母细胞的皮质中，除了在邻近纺锤体的皮质的环形带中。然而，在猪卵母细胞中没有观察到这样的IP 3 R1簇，并且其在体外和体内成熟卵母细胞之间的定位没有差异。接下来，我们试图澄清哪些因素调节IP 3 R1的定位，磷酸化和表达使用M期阶段依赖性激酶抑制剂。我们的研究结果表明，roscovitine（p34（cdc 2）激酶抑制剂）或U 0126（丝裂原活化蛋白激酶抑制剂）的治疗没有影响IP 3 R1的表达或定位在猪卵母细胞，虽然后者强烈抑制磷酸化。然而，用BI-2536（Polo样激酶1（Plk 1）的抑制剂）处理，以剂量依赖性方式显著降低猪卵母细胞中IP 3 R1的表达水平。这些结果表明，Plk 1参与了猪卵母细胞IP 3 R1表达的调控。
At fertilization, inositol 1,4,5-trisphosphate receptor type 1 (IP3 R1) has a crucial role in Ca(2+) release in mammals. Expression levels, localization and phosphorylation of IP3 R1 are important for its function, but it still remains unclear which molecule(s) regulates IP3 R1 behavior in pig oocytes. We examined whether there was a difference in localization of IP3 R1 after in vitro or in vivo maturation of pig oocytes. In mouse oocytes, large clusters of IP3 R1 were formed in the cortex of the oocyte except in a ring-shaped band of cortex adjacent to the spindle. However, no such clusters of IP3 R1 were observed in pig oocytes and there was no difference in its localization between in vitro and in vivo matured oocytes. We next tried to clarify which factor(s) regulates IP3 R1 localization, phosphorylation and expression using M-phase stage-dependent kinase inhibitors. Our results show that treatments with roscovitine (p34(cdc2) kinase inhibitor) or U0126 (mitogen-activated protein kinase inhibitor) did not affect IP3 R1 expression or localization in pig oocytes, although the latter strongly inhibited phosphorylation. However, treatment with BI-2536, an inhibitor of polo-like kinase 1 (Plk1), dramatically decreased the expression level of IP3 R1 in pig oocytes in a dose-dependent manner. From these results, it is suggested that Plk1 is involved in the regulation of IP3 R1 expression in pig oocytes.