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山羊卵母细胞孤雌激活和孤雌胚体外发育研究
许丹宁1, 田允波1, 黄运茂1, 陈学进2
0
(1.仲恺农业工程学院 生命科学学院,广州 510225;2.上海交通大学 医学院 实验动物科学部,上海 200025)
摘要:
为寻求适合于山羊卵母细胞孤雌激活的方法,本试验采用离子霉素对体外成熟山羊卵泡卵母细胞进行孤雌激活,同时比较了培养液中添加不同类型的血清对孤雌激活胚体外发育的影响。结果表明:用2.5、5.0和10.0 μmol/L 离子霉素处理卵母细胞,卵裂率分别为72.3%、75.6%和70.3%,囊胚率分别为40.8%、45.4%和39.4%,均无显著差异(P>0.05)。用5.0、10.0、15.0和20.0 μmol/L钙离子载体Ca-A23187处理卵母细胞,卵裂率分别为50.5%、62.5%、60.4%和60.2%,其中5.0 μmol/L Ca-A23187 处理组的卵裂率极显著低于其他处理组(P<0.01),但各组的囊胚率无显著差异(P<0.05)。用30、70、110和150 mL/L乙醇处理卵母细胞,卵裂率分别为40.6%、63.7%、64.2%和65.3%,其中30 mL/L乙醇组的卵裂率极显著低于其他各组(P<0.01),30和150 mL/L乙醇组的囊胚率极显著低于70和110 mL/L乙醇组(P<0.01)。在培养液与颗粒细胞共同培养条件下,分别添加100 mL/L的发情牛血清(OCS)、胎牛血清(FCS)和新生牛血清(NCS)。添加OCS和FCS后,孤雌胚的囊胚率分别为49.1%和48.2%,极显著高于添加NCS的(25.3%,P<0.01)。结果显示OCS和FBS能有效提高孤雌胚的体外发育能力,尤其是对提高孤雌胚的囊胚发育能力更佳。
关键词:  山羊  卵母细胞  体外成熟  孤雌激活  囊胚率
DOI:10.11841/j.issn.1007-4333.2010.05.016
投稿时间:2010-04-26
基金项目:农业部公益性行业(奶山羊)科研专项(3-45)第一作者:许丹宁,副教授,博士,主要从事兽医信息学及动物繁殖生物技术研究,E-mail:xdanning@126.com通讯作者:田允波,教授,博士,博士生导师,主要从事胚胎生物技术研究,E-mail:tyunbo@126.com
Study on parthenogenetic activated oocytes in goat andparthenogenetic embryos development in vitro
XU Dan-ning1, TIAN Yun-bo1, HUANG Yun-mao1, CHEN Xun-jin2
(1.College of Life Science,Zhongkai University of Agriculture and Engineering,Guangzhou 510225,China;2.School of Medicine,Department of Laboratory Animal Science,Shanghai Jiao Tong University,Shanghai 200025,China)
Abstract:
The present study was conducted to investigate activation of in vitro maturated goat follicular oocytes and parthenogenetic embryo development in vitro.The results showed that:The cleavage rate were 50.5%,62.5%,60.4% and 60.2% after activated by Ca-A23187 at 5.0,10.0,15.0 and 20.0 μmol/L respectively,and the cleavage rate of 5.0 μmol/ L was significantly lower than that of other groups (P<0.01).The cleavage rate were 40.6%,63.7%,64.2% and 65.3% after activated by ethanol at 30,70,110 and 150 mL/L respectively,and the cleavage rate of 30 mL/L ethanol was significantly lower than that of other groups (P<0.01).The blastocyst rate of 30,150 mL/L group was significantly lower than that of 70,110 mL/L group (P<0.01).When embryos were co-cultured with GCs in media with the same concentration (100 mL/L) of OCS,FCS and NCS,the blastocyst rate of parthenogenetic embryo added OCS and FCS (49.1%,48.2% resepctively) were significantly higher than which added NCS (25.3%,P<0.01).It was concluded that,adding OCS,FCS and NCS in media could significantly increased development of goat parthenogenetic embryos in vitro,moreover,the blastocyst rate of parthenogenetic embryos were significantly increased.
Key words:  goat  oocytes  in vitro maturation  parthenogenetic activation  blastocyst rate