猪不同直径卵泡内卵母细胞胞质成熟及体外发育潜力差异原因的初探

›› 2014, Vol. 41 ›› Issue (10): 165-171.

猪不同直径卵泡内卵母细胞胞质成熟及体外发育潜力差异原因的初探

焦明霞, 刘仲凤, 王峰, 刘世超, 格日乐其木格, 周洋, 孔庆然, 尹智, 郭诗萌, 刘忠华

东北农业大学生命学院, 黑龙江哈尔滨 150030

收稿日期:2014-03-04 出版日期:2014-10-20 发布日期:2014-10-29
通讯作者: 刘忠华 E-mail:liu086@126.com
作者简介:焦明霞(1988－),女,黑龙江人,硕士,研究方向:猪卵母细胞体外成熟及体外受精;刘仲凤(1985－),女,山东人,硕士,研究方向:猪卵母细胞体外成熟。
基金资助:
黑龙江省杰出青年基金(JC200905)

Preliminary Study on the Mechnism Account for the Difference of Cytoplasmic Maturation and in vitro Developmental Potential of Porcine Oocytes from Different Diameter Follicles

JIAO Ming-xia, LIU Zhong-feng, WANG Feng, LIU Shi-chao, Gerelchimeg, ZHOU Yang, KONG Qing-ran, YIN Zhi, GUO Shi-meng, LIU Zhong-hua

College of Life Science, Northeast Agricultural University, Harbin 150030, China

Received:2014-03-04 Online:2014-10-20 Published:2014-10-29

摘要/Abstract

摘要： 试验根据直径将猪卵泡分为2组:G1组(4~7 mm)和G2组(2~4 mm),对2组卵泡内获取的卵母细胞体外成熟率和发育潜能进行了比较,利用相对定量PCR检测了卵丘细胞中卵丘扩展相关基因Has2、Ptgs2、Ptx31及Pgr的表达水平,应用绝对定量PCR检测了成熟培养前后卵母细胞线粒体拷贝数,并利用5,5'-二巯基-2-硝基苯酸(DTNB)酶循环法检测了体外成熟培养过程中卵母细胞谷胱甘肽(GSH)的含量。结果显示,G1组和G2组卵母细胞体外成熟率分别为95.06%和68.19%,G1和G2组排出第一极体的成熟卵母细胞孤雌激活后的囊胚率分别为51.47%和29.44%,2组卵母细胞在体外成熟率和孤雌发育率上均差异显著(P<0.05)。G1组卵丘细胞在体外成熟培养过程中的扩展程度明显高于G2组,G1组卵母细胞对应的卵丘细胞扩展相关基因Has2、Ptgs2、Ptx31、Pgr的表达水平高于G2组(Has2基因在卵母细胞成熟培养0、24 h除外);G1组卵母细胞线粒体数、谷胱甘肽含量均高于G2组。以上结果表明,大卵泡来源的卵母细胞体外成熟能力和发育潜力优于小卵泡来源的卵母细胞,这可能与卵母细胞成熟过程中卵丘扩展程度、卵丘扩展相关基因表达激活情况、卵胞质内谷胱甘肽含量和线粒体拷贝数有关。

关键词: 猪卵母细胞; 体外成熟; 发育潜力; 卵泡直径; 卵丘细胞扩展相关基因; 谷胱甘肽; 线粒体

Abstract: We compared the in vitro maturation rate and parthenogenetic blastocyst rates of oocytes derived from follicles 4 to 7 mm (G1 group) and 2 to 4 mm (G2 group) in diameter. The cumulus expansion level,expansion relatived genes Has2,Ptgs2,Ptx31 and Pgr expression,GSH concentration and mitochondria copy number were also investigated. We found that the maturation rates of G1 and G2 were 95.06% and 68.19%,respectively,and the parthenogenetic blastocyste rates of these two groups were 51.47% and 29.44%,respectively,there were significant differences in maturation rates and partehnogenetic blastocysts rates between these two groups (P<0.05). The expansion level of cumulus cells in G1 was clearly higher than that in G2. The expression of cumulus cell expansion related gene Has2,Ptgs2,Ptx31 and Pgr in G1 group were higher than that of G2 group(except of Has2 gene in 0,24 h). Total GSH concentration and mitchondria numbers in G1 oocytes were higher than that in G2 oocytes. These results indicated that porcine oocytes from large follicles were better than these from small follicles in in vitro maturation and developmental potential,the cumulus cell expansion level,the expression of cumulus cell expansion related genes,concentration and mitochondria copy number might account for the difference.

Key words: porcine oocyte; in vitro maturation; developmental potential; follicle size; cumulus cell expansion related genes; GSH; mitochondrion

中图分类号:

S814

焦明霞, 刘仲凤, 王峰, 刘世超, 格日乐其木格, 周洋, 孔庆然, 尹智, 郭诗萌, 刘忠华. 猪不同直径卵泡内卵母细胞胞质成熟及体外发育潜力差异原因的初探[J]. , 2014, 41(10): 165-171.

JIAO Ming-xia, LIU Zhong-feng, WANG Feng, LIU Shi-chao, Gerelchimeg, ZHOU Yang, KONG Qing-ran, YIN Zhi, GUO Shi-meng, LIU Zhong-hua. Preliminary Study on the Mechnism Account for the Difference of Cytoplasmic Maturation and in vitro Developmental Potential of Porcine Oocytes from Different Diameter Follicles[J]. , 2014, 41(10): 165-171.

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