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褪黑素通过调节纺锤体组装检查点相关基因提高玻璃化冷冻-解冻小鼠卵母细胞的体外发育能力。

Melatonin Improves In Vitro Development of Vitrified-Warmed Mouse Germinal Vesicle Oocytes Potentially via Modulation of Spindle Assembly Checkpoint-Related Genes.

机构信息

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 & Histology, Shaheed Benazir Bhutto University of Veterinary and Animal Sciences, Sakrand 67210, Sindh, Pakistan.

出版信息

Cells. 2019 Aug 30;8(9):1009. doi: 10.3390/cells8091009.

Abstract

The present study aimed to investigate the effect of melatonin (MT) supplementation on in vitro maturation of vitrified mouse germinal vesicle (GV) oocytes. The fresh oocytes were randomly divided into three groups: untreated (control), or vitrified by open-pulled straw method without (vitrification group) or with MT supplementation (vitrification + MT group). After warming, oocytes were cultured in vitro, then the reactive oxygen species (ROS) and glutathione (GSH) levels, mitochondrial membrane potential, ATP levels, spindle morphology, mRNA expression of spindle assembly checkpoint (SAC)-related genes ), and their subsequent developmental potential in vitro were evaluated. The results showed that vitrification/warming procedures significantly decreased the percentage of GV oocytes developed to metaphase II (MII) stage, the mitochondrial membrane potential, ATP content, and GSH levels, remarkably increased the ROS levels, and significantly impaired the spindle morphology. The expressions of SAC-related genes were also altered in vitrified oocytes. However, when 10 mol/L MT was administered during the whole length of the experiment, the percentage of GV oocytes matured to MII stage was significantly increased, and the other indicators were also significantly improved and almost recovered to the normal levels relative to the control. Thus, we speculate that MT might regulate the mitochondrial membrane potential, ATP content, ROS, GSH, and expression of SAC-related genes, potentially increasing the in vitro maturation of vitrified-warmed mouse GV oocytes.

摘要

本研究旨在探讨褪黑素(MT)补充对玻璃化冷冻小鼠生发泡(GV)卵母细胞体外成熟的影响。新鲜卵母细胞随机分为三组:未经处理(对照组)、玻璃化冷冻(玻璃化组)或玻璃化冷冻并添加 MT(玻璃化+MT 组)。解冻后,将卵母细胞进行体外培养,然后评估活性氧(ROS)和谷胱甘肽(GSH)水平、线粒体膜电位、ATP 水平、纺锤体形态、纺锤体组装检查点(SAC)相关基因的 mRNA 表达及其随后的体外发育潜能。结果表明,玻璃化/解冻程序显著降低了GV 期卵母细胞发育为中期 II(MII)期的比例、线粒体膜电位、ATP 含量和 GSH 水平,显著增加了 ROS 水平,并显著损害了纺锤体形态。SAC 相关基因的表达也在玻璃化卵母细胞中发生改变。然而,当在整个实验过程中给予 10mol/L MT 时,GV 期卵母细胞成熟为 MII 期的比例显著增加,其他指标也显著改善,几乎恢复到对照的正常水平。因此,我们推测 MT 可能通过调节线粒体膜电位、ATP 含量、ROS、GSH 和 SAC 相关基因的表达,增加玻璃化冷冻-解冻小鼠 GV 卵母细胞的体外成熟。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4323/6770451/6d78e886c343/cells-08-01009-g001.jpg

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