Liang Shuang, Zhao Ming-Hui, Guo Jing, Choi Jeong-woo, Kim Nam-Hyung, Cui Xiang-Shun
Department of Animal Science, Chungbuk National University, Cheongju, Chungbuk, Republic of Korea.
Department of Animal Science, Chungbuk National University, Cheongju, Chungbuk, Republic of Korea.
Theriogenology. 2016 Mar 1;85(4):754-61.e1. doi: 10.1016/j.theriogenology.2015.10.021. Epub 2015 Oct 19.
PLK4, a polo-like kinase (PLK) family member that accumulates in the cytoplasm, has been identified as a crucial regulator of centriole formation. PLK4 also controls several essential cellular functions, including cytokinesis and gene expression. In this study, we investigated the expression and function of PLK4 during bovine oocyte meiotic maturation and subsequent embryo development. The PLK4 mRNA was detected in bovine oocytes at all developmental stages during meiotic maturation. Immunofluorescence staining showed that PLK4 protein exhibited a dynamic localization pattern in the oocyte cytoplasm during meiotic maturation, and fluorescence immunostaining markedly increased in metaphase II. When an interfering double-stranded RNA targeting PLK4 was injected into germinal vesicle-stage oocytes, PLK4 transcript levels decreased significantly in metaphase II oocytes (P < 0.05). The PLK4 knockdown caused spindle defects and chromosome misalignment and considerably reduced the amount of cortical and cytoplasmic actin. PLK4 was localized in the cytoplasm of early embryos, and PLK4 knockdown in germinal vesicle-stage oocytes led to failure in the early development of in vitro fertilized embryos (P < 0.05). Taken together, these results indicated that PLK4 plays crucial roles in bovine oocyte meiotic maturation and subsequent early embryo development.
PLK4是一种在细胞质中积累的类Polo样激酶(PLK)家族成员,已被确定为中心粒形成的关键调节因子。PLK4还控制着几种重要的细胞功能,包括胞质分裂和基因表达。在本研究中,我们调查了PLK4在牛卵母细胞减数分裂成熟及随后胚胎发育过程中的表达和功能。在减数分裂成熟的所有发育阶段的牛卵母细胞中均检测到PLK4 mRNA。免疫荧光染色显示,PLK4蛋白在减数分裂成熟过程中在卵母细胞细胞质中呈现动态定位模式,并且在中期II荧光免疫染色明显增加。当将靶向PLK4的干扰双链RNA注射到生发泡期卵母细胞中时,中期II卵母细胞中PLK4转录水平显著降低(P < 0.05)。PLK4敲低导致纺锤体缺陷和染色体排列紊乱,并显著减少皮质和细胞质肌动蛋白的量。PLK4定位于早期胚胎的细胞质中,并且生发泡期卵母细胞中PLK4敲低导致体外受精胚胎早期发育失败(P < 0.05)。综上所述,这些结果表明PLK4在牛卵母细胞减数分裂成熟及随后的早期胚胎发育中起关键作用。