牦牛卵母细胞及体外受精胚胎早期发育基因差异表达的研究

doi:10.11843/j.issn.0366-6964.2013.10.010

畜牧兽医学报 ›› 2013, Vol. 44 ›› Issue (10): 1576-1581.doi: 10.11843/j.issn.0366-6964.2013.10.010

牦牛卵母细胞及体外受精胚胎早期发育基因差异表达的研究

郭宪，丁学智，王宏博，包鹏甲，阎萍^*

(中国农业科学院兰州畜牧与兽药研究所，甘肃省牦牛繁育工程重点实验室, 兰州 730050）

收稿日期:2013-01-05 出版日期:2013-10-23 发布日期:2013-10-28
通讯作者: 阎萍，E-mail: pingyan@sohu.com
作者简介:郭宪（1978-），男，甘肃环县人，博士，副研究员，主要从事动物遗传育种与繁殖研究，E-mail: guoxian2006@sina.com
基金资助:
中央级公益性科研院所基本科研业务费专项（1610322011002）；现代农业（肉牛牦牛）产业技术体系建设专项（CARS-38）；甘肃省科技重大专项（1102NKDA027）

Gene Expression of Yak Oocytes and Preimplantation Embryos Derived from in vitro Fertilization

GUO Xian, DING Xue-zhi, WANG Hong-bo, BAO Peng-jia, YAN Ping^*

(Key Laboratory of Yak Breeding Engineering of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China)

Received:2013-01-05 Online:2013-10-23 Published:2013-10-28

摘要/Abstract

摘要：

旨在了解牦牛卵母细胞、早期胚胎不同发育阶段相关基因mRNA表达规律，为早期胚胎发育基因表达的调控机制提供分子依据。应用mRNA差异显示技术选取表达量存在差异的基因条带，进行测序和同源性分析，并利用RT-PCR技术检测差异基因在卵母细胞、2-细胞、8-细胞和囊胚中的相对表达水平。结果表明：4条差异条带中，除桑椹胚期1个基因在GenBank中未检测到同源性外，其余3个基因分别与锌指MYND11（ZMYND11）、核小体组装蛋白1样1（NAP1L1）、核糖体蛋白L27（RPL27A）高度同源。ZMYND11基因在成熟卵母细胞、2-细胞、8-细胞、囊胚阶段其表达量呈上升趋势，但无显著性差异。NAP1L1基因在成熟卵母细胞、2-细胞、囊胚阶段其表达量相对较低，而在8-细胞表达量急剧升高（P<0.05）。RPL27A基因从成熟卵母细胞经2-细胞到8-细胞随胚胎发育进程表达量逐渐降低，到囊胚期表达量显著升高（P<0.05）。基因ZMYND11、RPL27A和NAP1L1在牦牛卵母细胞和早期胚胎发育过程中mRNA表达量存在时间性差异，可能与早期胚胎的正常发育及不同的生理活动有关。

关键词: 牦牛, 早期胚胎, 胚胎发育, 基因表达

Abstract:

The purpose of this study was to clarify differential pattern of mRNA expression for genes related to different developmental stages of yak oocytes and preimplatation embryos, and to provide molecular evidence for gene expression of preimplatation embryonic development. The mRNA differential display reverse transcription PCR was used to identify differential expression bands for sequencing and alignment. The levels of mRNA relative expression were evaluated by RT-PCR in oocyte, 2-cell embryo, 8-cell embryo and blastocyst in yak. The results showed that three differential expression bands were highly homologous with zinc finger MYND-type containing 11 (ZMYND11), ribosomal protein L27a (RPL27A) and nucleosome assembly protein 1-like 1 (NAP1L1), respectively, except that one gene during morula stage showed no homology to any other known genes in GenBank. The relative expression levels of ZMYND11 genes increased in matured oocyte, 2-cell embryo, 8-cell embryo and in the blastocyst, with no significant differences between these stages (P>0.05). The relative expression levels of NAP1L1 genes were relatively low in matured oocyte, 2-cell embryo and blastocyst, with sharp increase in 8-cell embryo (P<0.05). The relative expression levels of RPL27A genes decreased during development, from matured oocyte to 2-cell, then to 8-cell embryo, and increased significantly in the blastocyst (P<0.05). ZMYND11, RPL27A and NAP1L1, the mRNA expression levels of which showed differences in different development stages of yak oocytes and preimplantation embryos, might be related to normal development and different physiological activities of preimplantation embryo in yak.

Key words: yak, preimplantation embryo, embryo development, gene expression

中图分类号:

郭宪，丁学智，王宏博，包鹏甲，阎萍. 牦牛卵母细胞及体外受精胚胎早期发育基因差异表达的研究[J]. 畜牧兽医学报, 2013, 44(10): 1576-1581.

GUO Xian, DING Xue-zhi, WANG Hong-bo, BAO Peng-jia, YAN Ping. Gene Expression of Yak Oocytes and Preimplantation Embryos Derived from in vitro Fertilization[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(10): 1576-1581.

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牦牛卵母细胞及体外受精胚胎早期发育基因差异表达的研究

Gene Expression of Yak Oocytes and Preimplantation Embryos Derived from in vitro Fertilization

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