Department of Histology and Embryology, Akdeniz University School of Medicine, Antalya, Turkey.
Department of Histology and Embryology, Ankara University School of Medicine, Ankara, Turkey.
J Cell Biochem. 2019 Mar;120(3):4694-4709. doi: 10.1002/jcb.27759. Epub 2018 Oct 9.
The embryonic poly(A)-binding protein (EPAB) functions in the translational regulation of the maternal messenger RNAs (mRNAs) required during oocyte maturation, fertilization, and early embryo development. Since there is no antibody specific to mammalian EPAB protein, all studies related to the Epab gene could be performed at the mRNA levels except for the investigations in the Xenopus. In this study, we have produced an EPAB-specific antibody. When we examined its expressional distribution in the mouse gonadal and somatic tissues, the EPAB protein was found to be expressed only in the mouse ovary and testis tissues, but it is undetectable level in the somatic tissues including stomach, liver, heart, small intestine, and kidney. Additionally, the spatial and temporal expression patterns of the EPAB and poly(A)-binding protein cytoplasmic 1 (PABPC1) proteins were analyzed in the mouse germinal vesicle (GV) and metaphase II (MII) oocytes, one-cell, and two-cell embryos. While EPAB expression gradually decreased from GV oocytes to two-cell embryos, the PABPC1 protein level progressively increased from GV oocytes to one-cell embryos and remarkably declined in the two-cell embryos ( P < 0.05). We have also described herein that the EPAB protein interacted with Epab, Pabpc1, Ccnb1, Gdf9, and Bmp15 mRNAs dependent upon the developmental stages of the mouse oocytes and early embryos. As a result, we have first produced an EPAB-specific antibody and characterized its expression patterns and interacting mRNAs in the mouse oocytes and early embryos. The findings suggest that EPAB in cooperation with PABPC1 implicate in the translational control of maternal mRNAs during oogenesis and early embryo development.
胚胎多聚腺苷酸结合蛋白 (EPAB) 参与卵母细胞成熟、受精和早期胚胎发育过程中所需的母体信使 RNA(mRNA) 的翻译调控。由于没有针对哺乳动物 EPAB 蛋白的特异性抗体，除了在非洲爪蟾中的研究外，所有与 Epab 基因相关的研究都可以在 mRNA 水平上进行。在本研究中，我们制备了一种 EPAB 特异性抗体。当我们检测其在小鼠性腺和体细胞组织中的表达分布时，发现 EPAB 蛋白仅在小鼠卵巢和睾丸组织中表达，但在包括胃、肝、心、小肠和肾在内的体细胞组织中检测不到。此外，还分析了 EPAB 和多聚腺苷酸结合蛋白细胞质 1 (PABPC1) 蛋白在小鼠生发泡期 (GV) 和中期 II (MII) 卵母细胞、单细胞和二细胞胚胎中的时空表达模式。虽然 EPAB 表达从 GV 卵母细胞逐渐降低到二细胞胚胎，但 PABPC1 蛋白水平从 GV 卵母细胞逐渐增加到单细胞胚胎，并在二细胞胚胎中显著降低 (P<0.05)。我们还描述了 EPAB 蛋白与 Epab、Pabpc1、Ccnb1、Gdf9 和 Bmp15 mRNA 的相互作用，这种相互作用依赖于小鼠卵母细胞和早期胚胎的发育阶段。因此，我们首次制备了一种 EPAB 特异性抗体，并在小鼠卵母细胞和早期胚胎中对其表达模式和相互作用的 mRNA 进行了特征描述。研究结果表明，EPAB 与 PABPC1 一起参与了卵子发生和早期胚胎发育过程中母体 mRNA 的翻译调控。
