The Research Centre for Laboratory Animal Science, State Key Laboratory of Reproductive Regulation & Breeding of Grassland Livestock, School of Life Science, Inner Mongolia University , Hohhot , People's Republic of China.
Department of Obstetrics and Gynecology, Inner Mongolia People's Hospital , Hohhot , People's Republic of China.
Cell Cycle. 2019 Oct;18(20):2784-2799. doi: 10.1080/15384101.2019.1661173. Epub 2019 Sep 3.
Aneuploidy caused by abnormal chromosome segregation during early embryo development leads to embryonic death or congenital malformation. Centromere protein F (CENPF) is a member of centromere protein family that regulates chromosome segregation during mitosis. However, its necessity in early embryo development has not been fully investigated. In this study, expression and function of CENPF was investigated in mouse early embryogenesis. Detection of CENPF expression and localization revealed a cytoplasm, spindle and nuclear membrane related dynamic pattern throughout mitotic progression. Farnesyltransferase inhibitor (FTI) was employed to inhibit CENPF farnesylation in zygotes. The results showed that CENPF degradation was inhibited and its specific localization on nuclear membranes in morula and blastocyst vanished after FTI treatment. Also, CAAX motif mutation leads to failure of CENPF-C630 localization in morula and blastocyst. These results indicate that farnesylation plays a key role during CENPF degradation and localization in early embryos. To further assess CENPF function in parthenogenetic or fertilized embryos development, morpholino (MO) and Trim-Away were used to disturb CENPF function. CENPF knockdown in Metaphase II (MII) oocytes, zygotes or embryos with MO approach resulted in failure to develop into morulae and blastocysts, revealing its indispensable role in both parthenogenetic and fertilized embryos. Disturbing of CENPF with Trim-Away approach in zygotes resulted in impaired development of 2-cell and 4-cell, but did not affect the morula and blastocyst formation because of the recovered expression of CENPF. Taken together, our data suggest CENPF plays an important role during early embryonic development in mice. : CENPF: centromere protein F; MO: morpholino; FTI: Farnesyltransferase inhibitor; CENPE: centromere protein E; IVF: fertilization; MII: metaphase II; SAC: spindle assembly checkpoint; Mad1: mitotic arrest deficient 1; BUB1: budding uninhibited by benzimidazole 1; BUBR1: BUB1 mitotic checkpoint serine/threonine kinase B; Cdc20: cell division cycle 20.
早期胚胎发育过程中染色体分离异常导致的非整倍体可导致胚胎死亡或先天性畸形。着丝粒蛋白 F（CENPF）是着丝粒蛋白家族的成员，可调节有丝分裂过程中的染色体分离。然而，其在早期胚胎发育中的必要性尚未得到充分研究。在这项研究中，研究了 CENPF 在小鼠早期胚胎发生中的表达和功能。通过检测 CENPF 的表达和定位，揭示了整个有丝分裂进程中细胞质、纺锤体和核膜相关的动态模式。法呢基转移酶抑制剂（FTI）被用于抑制受精卵中的 CENPF 法尼基化。结果表明，FTI 处理后，CENPF 降解受到抑制，其在桑葚胚和囊胚中的核膜上的特异性定位消失。此外，CAAX 基序突变导致 CENPF-C630 在桑葚胚和囊胚中的定位失败。这些结果表明，法尼基化在 CENPF 降解和早期胚胎中的定位中起关键作用。为了进一步评估 CENPF 在孤雌激活或受精胚胎发育中的功能，使用了 morpholino（MO）和 Trim-Away 来干扰 CENPF 功能。用 MO 方法干扰 MII 期卵母细胞、受精卵或胚胎中的 CENPF 功能导致其无法发育成桑葚胚和囊胚，表明其在孤雌激活和受精胚胎中都是不可或缺的。用 Trim-Away 方法干扰受精卵中的 CENPF 导致 2-细胞和 4-细胞的发育受损，但不影响桑葚胚和囊胚的形成，因为 CENPF 的表达得到了恢复。总之，我们的数据表明 CENPF 在小鼠早期胚胎发育中发挥着重要作用。
