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在第一次减数分裂期间,抑制类Polo样激酶1会导致猪卵母细胞中的染色体排列不齐和纺锤体异常。

Polo-like kinase 1 inhibition results in misaligned chromosomes and aberrant spindles in porcine oocytes during the first meiotic division.

作者信息

Liao Y, Lin D, Cui P, Abbasi B, Chen C, Zhang Z, Zhang Y, Dong Y, Rui R, Ju S

机构信息

College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.

出版信息

Reprod Domest Anim. 2018 Feb;53(1):256-265. doi: 10.1111/rda.13102. Epub 2017 Nov 16.

Abstract

Polo-like kinase 1 (Plk1), a type of serine/threonine protein kinase, has been implicated in various functions in the regulation of mitotic processes. However, these kinase's roles in meiotic division are not fully understood, particularly in the meiotic maturation of porcine oocytes. In this study, the expression and spatiotemporal localization of Plk1 were initially assessed in the meiotic process of pig oocytes by utilizing Western blotting with immunofluorescent staining combined with confocal microscopy imaging technique. The results showed that Plk1 was expressed and exhibited a dynamic subcellular localization throughout the meiotic process. After germinal vesicle breakdown (GVBD), Plk1 was detected prominently around the condensed chromosomes and subsequently exhibited a similar subcellular localization to α-tubulin throughout subsequent meiotic phases, with particular enrichment being observed near spindle poles at MI and MII. Inhibition of Plk1 via a highly selective inhibitor, GSK461364, led to the failure of first polar body extrusion in porcine oocytes, with the majority of the treated oocytes being arrested in GVBD. Further subcellular structure examination results indicated that Plk1 inhibition caused the great majority of oocytes with spindle abnormalities and chromosome misalignment during the first meiotic division. The results of this study illustrate that Plk1 is critical for the first meiotic division in porcine oocytes through its influence on spindle organization and chromosome alignment, which further affects the ensuing meiotic cell cycle progression.

摘要

Polo样激酶1(Plk1)是一种丝氨酸/苏氨酸蛋白激酶,参与有丝分裂过程调控的多种功能。然而,该激酶在减数分裂中的作用尚未完全明确,尤其是在猪卵母细胞的减数分裂成熟过程中。在本研究中,通过免疫荧光染色结合共聚焦显微镜成像技术的蛋白质免疫印迹法,初步评估了Plk1在猪卵母细胞减数分裂过程中的表达及时空定位。结果显示,Plk1在整个减数分裂过程中均有表达,并呈现动态的亚细胞定位。生发泡破裂(GVBD)后,在浓缩染色体周围可明显检测到Plk1,随后在后续减数分裂阶段其亚细胞定位与α-微管蛋白相似,在减数第一次分裂(MI)和减数第二次分裂(MII)的纺锤体极附近观察到其特别富集。通过高度选择性抑制剂GSK461364抑制Plk1,导致猪卵母细胞第一极体排出失败,大多数处理后的卵母细胞停滞在GVBD阶段。进一步的亚细胞结构检查结果表明,抑制Plk1会导致大多数卵母细胞在第一次减数分裂期间出现纺锤体异常和染色体排列紊乱。本研究结果表明,Plk1通过影响纺锤体组织和染色体排列,对猪卵母细胞的第一次减数分裂至关重要,进而影响随后的减数分裂细胞周期进程。

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