Pan Bo, Qazi Izhar Hyder, Guo Shichao, Yang Jingyu, Qin Jianpeng, Lv Tianyi, Zang Shengqin, Zhang Yan, Zeng Changjun, Meng Qingyong, Han Hongbing, Zhou Guangbin
Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, 611130, China.
Department of Veterinary Anatomy and Histology, Shaheed Benazir Bhutto University of Veterinary and Animal Sciences, Sakrand, Sindh, 67210, Pakistan.
J Anim Sci Biotechnol. 2021 Jul 16;12(1):84. doi: 10.1186/s40104-021-00605-y.
This study investigated the effect of melatonin (MT) on cell cycle (G1/S/G2/M) of parthenogenetic zygotes developed from vitrified-warmed mouse metaphase II (MII) oocytes and elucidated the potential mechanism of MT action in the first cleavage of embryos.
After vitrification and warming, oocytes were parthenogenetically activated (PA) and in vitro cultured (IVC). Then the spindle morphology and chromosome segregation in oocytes, the maternal mRNA levels of genes including Miss, Doc1r, Setd2 and Ythdf2 in activated oocytes, pronuclear formation, the S phase duration in zygotes, mitochondrial function at G1 phase, reactive oxygen species (ROS) level at S phase, DNA damage at G2 phase, early apoptosis in 2-cell embryos, cleavage and blastocyst formation rates were evaluated. The results indicated that the vitrification/warming procedures led to following perturbations 1) spindle abnormalities and chromosome misalignment, alteration of maternal mRNAs and delay in pronucleus formation, 2) decreased mitochondrial membrane potential (MMP) and lower adenosine triphosphate (ATP) levels, increased ROS production and DNA damage, G1/S and S/G2 phase transition delay, and delayed first cleavage, and 3) increased early apoptosis and lower levels of cleavage and blastocyst formation. Our results further revealed that such negative impacts of oocyte cryopreservation could be alleviated by supplementation of warming, recovery, PA and IVC media with 10 mol/L MT before the embryos moved into the 2-cell stage of development.
MT might promote cell cycle progression via regulation of MMP, ATP, ROS and maternal mRNA levels, potentially increasing the first cleavage of parthenogenetic zygotes developed from vitrified-warmed mouse oocytes and their subsequent development.
本研究调查了褪黑素(MT)对玻璃化冷冻复苏的小鼠中期Ⅱ(MII)卵母细胞孤雌激活合子细胞周期(G1/S/G2/M)的影响,并阐明了MT在胚胎第一次卵裂中的潜在作用机制。
玻璃化冷冻复苏后,卵母细胞进行孤雌激活(PA)并体外培养(IVC)。然后评估卵母细胞中的纺锤体形态和染色体分离、激活卵母细胞中包括Miss、Doc1r、Setd2和Ythdf2等基因的母源mRNA水平、原核形成、合子中S期持续时间、G1期线粒体功能、S期活性氧(ROS)水平、G2期DNA损伤、2细胞胚胎早期凋亡、卵裂率和囊胚形成率。结果表明,玻璃化冷冻复苏过程导致了以下扰动:1)纺锤体异常和染色体排列紊乱、母源mRNA改变以及原核形成延迟;2)线粒体膜电位(MMP)降低和三磷酸腺苷(ATP)水平降低、ROS产生增加和DNA损伤、G1/S和S/G2期转换延迟以及第一次卵裂延迟;3)早期凋亡增加以及卵裂率和囊胚形成率降低。我们的结果进一步表明,在胚胎进入2细胞发育阶段之前,在玻璃化冷冻复苏、恢复、PA和IVC培养基中添加10μmol/L MT可以减轻卵母细胞冷冻保存的这些负面影响。
MT可能通过调节MMP、ATP、ROS和母源mRNA水平促进细胞周期进程,潜在地增加玻璃化冷冻复苏的小鼠卵母细胞孤雌激活合子的第一次卵裂及其后续发育。