Kim Kyeoung-Hwa, Kim Eun-Young, Lee Kyung-Ah
Graduate School of Life Science and Biotechnology, Pochon CHA University College of Medicine, CHA Research Institute, Fertility Center, CHA General Hospital, Seoul 135-081, Korea.
Biol Reprod. 2008 Dec;79(6):1192-201. doi: 10.1095/biolreprod.108.068478. Epub 2008 Aug 27.
Previously, we found high levels of skin-embryo-brain-oocyte homeobox (Sebox) gene expression in germinal vesicle (GV)-stage oocytes. The objective of the present study was to determine the role played by SEBOX in oocyte maturation and early embryogenesis using RNA interference (RNAi). Microinjection of Sebox double-stranded RNA into GV oocytes resulted in a marked decrease in Sebox mRNA and protein expression. However, Sebox RNAi affects neither oocyte maturation rate nor morphological characteristics, including spindle and chromosomal organization of metaphase II oocytes. In addition, Sebox RNAi had no discernible effect on the activities of M-phase promoting factor or mitogen-activated protein kinase. In contrast, microinjection of Sebox double-stranded RNA into pronuclear-stage embryos resulted in holding embryo development at the two-cell (84.9%) and the four- and eight-cell (15.1%) stages. We concluded that Sebox is a new addition to maternal effect genes that produced and stored in oocytes and function in preimplantation embryo development.
此前,我们发现生发泡(GV)期卵母细胞中皮肤-胚胎-脑-卵母细胞同源框(Sebox)基因表达水平很高。本研究的目的是利用RNA干扰(RNAi)来确定SEBOX在卵母细胞成熟和早期胚胎发育中所起的作用。将Sebox双链RNA显微注射到GV期卵母细胞中,导致Sebox mRNA和蛋白表达显著降低。然而,Sebox RNAi既不影响卵母细胞成熟率,也不影响形态特征,包括中期II卵母细胞的纺锤体和染色体组织。此外,Sebox RNAi对M期促进因子或丝裂原活化蛋白激酶的活性没有明显影响。相反,将Sebox双链RNA显微注射到原核期胚胎中,导致胚胎发育停滞在二细胞期(84.9%)以及四细胞期和八细胞期(15.1%)。我们得出结论,Sebox是母源效应基因中的一个新成员,这些基因在卵母细胞中产生和储存,并在植入前胚胎发育中发挥作用。