Moon Jeonghyeon, Roh Sangho
Cellular Reprogramming and Embryo Biotechnology Laboratory, Dental Research Institute, BK21 PLUS Dental Life Science, Seoul National University School of Dentistry, Seoul 08826, Korea.
J Vet Sci. 2019 Jan 31;20(1):2-9. doi: 10.4142/jvs.2019.20.1.2.
Somatic cell nuclear transfer (SCNT) has various applications in research, as well as in the medical field and animal husbandry. However, the efficiency of SCNT is low and the accurate mechanism of SCNT in murine embryo development is unreported. In general, the developmental rate of SCNT murine embryos is lower than counterparts. In previous studies, polo-like kinase 1 (Plk1) was reported to be a crucial element in cell division including centrosome maturation, cytokinesis, and spindle formation. In an initial series of experiments in this study, BI2536, a Plk1 inhibitor, was treated to -fertilized embryos and the embryos failed to develop beyond the 2-cell stage. This confirmed previous findings that Plk1 is crucial for the first mitotic division of murine embryos. Next, we investigated Plk1's localization and intensity by immunofluorescence analysis. In contrast to normally developed embryos, SCNT murine embryos that failed to develop exhibited two types of Plk1 expressions; a low Plk1 expression pattern and ectopic expression of Plk1. The results show that Plk1 has a critical role in SCNT murine embryos. In conclusion, this study demonstrated that the SCNT murine embryos fail to develop beyond the 2-cell stage, and the embryos show abnormal Plk1 expression patterns, which may one of the main causes of developmental failure of early SCNT murine embryos.
体细胞核移植(SCNT)在研究、医学领域和畜牧业中有多种应用。然而,SCNT的效率较低,且SCNT在小鼠胚胎发育中的准确机制尚未见报道。一般来说,SCNT小鼠胚胎的发育率低于对照胚胎。在以往的研究中,据报道,polo样激酶1(Plk1)是细胞分裂中的关键因素,包括中心体成熟、胞质分裂和纺锤体形成。在本研究的一系列初步实验中,将Plk1抑制剂BI2536处理于受精后的胚胎,这些胚胎未能发育至2细胞期以后。这证实了先前的研究结果,即Plk1对小鼠胚胎的第一次有丝分裂至关重要。接下来,我们通过免疫荧光分析研究了Plk1的定位和强度。与正常发育的胚胎相比,未能发育的SCNT小鼠胚胎表现出两种类型的Plk1表达;低Plk-1表达模式和Plk1的异位表达。结果表明,Plk1在SCNT小鼠胚胎中起关键作用。总之,本研究表明,SCNT小鼠胚胎未能发育至2细胞期以后,且胚胎表现出异常的Plk1表达模式,这可能是早期SCNT小鼠胚胎发育失败的主要原因之一。