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酪蛋白激酶2调节纺锤体组装检查点以协调猪卵母细胞减数分裂进程。

Casein kinase 2 modulates the spindle assembly checkpoint to orchestrate porcine oocyte meiotic progression.

作者信息

ShiYang Xiayan, Miao Yilong, Cui Zhaokang, Lu Yajuan, Zhou Changyin, Zhang Yu, Xiong Bo

机构信息

College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095 China.

出版信息

J Anim Sci Biotechnol. 2020 Apr 8;11:31. doi: 10.1186/s40104-020-00438-1. eCollection 2020.

Abstract

BACKGROUND

CK2 (casein kinase 2) is a serine/threonine-selective protein kinase that has been involved in a variety of cellular processes such as DNA repair, cell cycle control and circadian rhythm regulation. However, its functional roles in oocyte meiosis have not been fully determined.

RESULTS

We report that CK2 is essential for porcine oocyte meiotic maturation by regulating spindle assembly checkpoint (SAC). Immunostaining and immunoblotting analysis showed that CK2 was constantly expressed and located on the chromosomes during the entire oocyte meiotic maturation. Inhibition of CK2 activity by its selective inhibitor CX-4945 impaired the first polar body extrusion and arrested oocytes at M I stage, accompanied by the presence of BubR1 at kinetochores, indicative of activated SAC. In addition, we found that spindle/chromosome structure was disrupted in CK2-inhibited oocytes due to the weakened microtubule stability, which is a major cause resulting in the activation of SAC. Last, we found that the level DNA damage as assessed by γH2A.X staining was considerably elevated when CK2 was inhibited, suggesting that DNA damage might be another critical factor leading to the SAC activation and meiotic failure of oocytes.

CONCLUSIONS

Our findings demonstrate that CK2 promotes the porcine oocyte maturation by ensuring normal spindle assembly and DNA damage repair.

摘要

背景

酪蛋白激酶2(CK2)是一种丝氨酸/苏氨酸选择性蛋白激酶,参与多种细胞过程,如DNA修复、细胞周期调控和昼夜节律调节。然而,其在卵母细胞减数分裂中的功能作用尚未完全确定。

结果

我们报道CK2通过调节纺锤体组装检查点(SAC)对猪卵母细胞减数分裂成熟至关重要。免疫染色和免疫印迹分析表明,在整个卵母细胞减数分裂成熟过程中,CK2持续表达并定位于染色体上。其选择性抑制剂CX-4945抑制CK2活性会损害第一极体排出,并使卵母细胞停滞在MI期,同时动粒上存在BubR1,表明SAC被激活。此外,我们发现由于微管稳定性减弱,CK2抑制的卵母细胞中纺锤体/染色体结构被破坏,这是导致SAC激活的主要原因。最后,我们发现当CK2被抑制时,通过γH2A.X染色评估的DNA损伤水平显著升高,表明DNA损伤可能是导致卵母细胞SAC激活和减数分裂失败的另一个关键因素。

结论

我们的研究结果表明,CK2通过确保正常的纺锤体组装和DNA损伤修复来促进猪卵母细胞成熟。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fbbb/7140493/a0b2b5ad57b6/40104_2020_438_Fig3_HTML.jpg

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