褪黑素对牛卵母细胞体外成熟及孤雌胚胎发育的影响

发布时间：2018-10-19 06:35

【摘要】：褪黑素对生殖系统发挥着重要调节作用,本研究旨在探究褪黑素对牛卵母细胞体外成熟和孤雌胚胎发育的影响及作用机制。本试验用化学激活法对牛卵母细胞孤雌激活,且在卵母细胞体外成熟液和胚胎体外培养液中添加不同浓度(0 pmol/ml、10 pmol/ml、20 pmol/ml、30 pmol/ml)的褪黑素,以检测牛卵母细胞成熟率和孤雌胚胎发育效果,同时利用DCHF-DA染色方法检测卵母细胞中ROS水平,采用免疫荧光技术检测褪黑素受体MT1在牛早期胚胎发育过程中的表达。结果表明,外源褪黑素可促进卵母细胞体外成熟,卵母细胞成熟率随褪黑素浓度的增加而升高。褪黑素显著降低了卵母细胞中的ROS水平(P0.05),且卵母细胞中的ROS水平随着褪黑素浓度增加而降低。褪黑素可显著提高孤雌胚胎的卵裂率、桑葚胚率和囊胚率(P0.05),促进胚胎发育。相比对照组,10pmol/ml褪黑素剂量更有利于胚胎发育,桑葚胚率和囊胚率都显著高于其他试验组。另外,本研究首次探究了褪黑素受体MT1在早期胚胎发育过程中的表达分布。MT1在未卵裂的激活卵母细胞中主要分布于细胞膜上,随着孤雌胚胎卵裂的进行,受体表达量逐渐增加,主要表达定位于细胞膜上,随着胚胎发育的进行,MT1在卵裂球内部开始表达和分布,退化胚胎中MT1表达水平较低。可见,褪黑素可促进卵母细胞成熟和胚胎发育,且褪黑素受体MT1在胚胎发育早期已开始表达。
[Abstract]:Melatonin plays an important role in regulating the reproductive system. The aim of this study was to investigate the effects of melatonin on bovine oocytes maturation in vitro and parthenogenetic embryo development. In this experiment, bovine oocyte parthenogenetic activation was carried out by chemical activation, and melatonin (0 pmol/ml,10 pmol/ml,20 pmol/ml,30 pmol/ml) was added to oocyte maturation fluid and embryo culture medium in order to detect the maturation rate of bovine oocyte and the effect of parthenogenetic embryo development. At the same time, the level of ROS in oocytes was detected by DCHF-DA staining, and the expression of melatonin receptor MT1 in bovine early embryo development was detected by immunofluorescence technique. The results showed that exogenous melatonin could promote the maturation of oocytes in vitro, and the maturation rate of oocytes increased with the increase of melatonin concentration. Melatonin significantly decreased the level of ROS in oocytes (P0.05), and the level of ROS in oocytes decreased with the increase of melatonin concentration. Melatonin could significantly increase the cleavage rate, morula rate and blastocyst rate of parthenogenetic embryos (P0.05). Compared with the control group, the dose of 10pmol/ml melatonin was more favorable for embryo development, and the morula rate and blastocyst rate were significantly higher than those in other experimental groups. In addition, the expression and distribution of melatonin receptor MT1 during early embryonic development were investigated for the first time. The expression of melatonin receptor MT1 increased gradually with the cleavage of parthenogenetic embryos, and MT1 was mainly distributed on the membrane of activated oocytes without cleavage. The expression of MT1 was mainly located on the cell membrane. With the development of the embryo, MT1 began to express and distribute in the blastomere, and the expression level of MT1 was lower in the degenerated embryos. Melatonin can promote oocyte maturation and embryo development, and the expression of melatonin receptor MT1 has already begun in the early stage of embryonic development.
【作者单位】：安徽科技学院动物科学学院;华中农业大学动物科学技术学院;
【基金】：国家自然科学基金项目(31301972) 安徽省自然科学基金项目(1308085QC66) 安徽省教育厅重点项目(KJ2009B090) 现代农业产业技术体系建设专项(CARS-37-04B)
【分类号】：S823

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