Department of Histology and Embryology, School of Medicine, Akdeniz University, Campus, 07070, Antalya, Turkey.
J Assist Reprod Genet. 2018 Mar;35(3):503-513. doi: 10.1007/s10815-017-1087-z. Epub 2017 Nov 22.
DNA methylation is an epigenetic mechanism that plays critical roles during mammalian oocyte and preimplantation embryo development. It is achieved by adding a methyl group to the fifth carbon atom of cytosine residues within cytosine-phosphate-guanine (CpG) and non-CpG dinucleotide sites using DNA methyltransferase (DNMT) enzymes for de novo and maintenance methylation processes. DNMT1, DNMT3A, and DNMT3B play important roles in establishing methylation of developmentally related genes in oocytes and early embryos. The purpose of this study is to identify the effect of superovulation on the expression and subcellular localizations of these three DNMT enzymes in the mouse oocytes and early embryos.
Three groups composed of control, normal dose [5 IU pregnant mare serum gonadotropin (PMSG) and 5 IU human chorionic gonadotropin (hCG)], and high dose [7.5 IU PMSG and 7.5 IU hCG] were created from 4-5-week-old female BALB/c mice. The relative expression and subcellular localizations of the DNMT proteins in the control and experiment groups have been characterized by using immunofluorescence staining subsequently analyzed in detailed.
DNMT1, DNMT3A, and DNMT3B protein expression in the germinal vesicle and metaphase II oocytes and in one-cell and two-cell embryos differed significantly when some of the normal- and high-dose groups were compared with the control counterparts.
This study has demonstrated for the first time that superovulation alters expression levels of the DNMT proteins, a finding that indicates that certain developmental defects in superovulated oocytes and early embryos may result from impaired DNA methylation processes.
DNA 甲基化是一种表观遗传机制,在哺乳动物卵母细胞和着床前胚胎发育过程中发挥着关键作用。它是通过 DNA 甲基转移酶(DNMT)酶将甲基添加到胞嘧啶残基的第五个碳原子上,在胞嘧啶-磷酸-鸟嘌呤(CpG)和非 CpG 二核苷酸位点上实现的,用于从头和维持甲基化过程。DNMT1、DNMT3A 和 DNMT3B 在卵母细胞和早期胚胎中发育相关基因的甲基化建立中发挥重要作用。本研究旨在确定超排卵对这些三种 DNMT 酶在小鼠卵母细胞和早期胚胎中的表达和亚细胞定位的影响。
从 4-5 周龄雌性 BALB/c 小鼠中创建了三组:对照组、正常剂量组[5 IU 孕马血清促性腺激素(PMSG)和 5 IU 人绒毛膜促性腺激素(hCG)]和高剂量组[7.5 IU PMSG 和 7.5 IU hCG]。随后使用免疫荧光染色对控制组和实验组中 DNMT 蛋白的相对表达和亚细胞定位进行了特征描述,并进行了详细分析。
在一些正常剂量和高剂量组与对照组相比,DNMT1、DNMT3A 和 DNMT3B 蛋白在生发泡和中期卵母细胞以及一细胞和两细胞胚胎中的表达存在显著差异。
本研究首次证明超排卵改变了 DNMT 蛋白的表达水平,这表明超排卵卵母细胞和早期胚胎中的某些发育缺陷可能是由于 DNA 甲基化过程受损所致。