乙二胺四乙酸钠、能量基质、氨基酸对猪卵母细胞体外成熟和早期孤雌发育的影响  

Effects of ethylenediaminetetraacetic acid,energy substrate,amino acids on in vitro maturation of porcine oocytes and in vitro development of porcine parthenogenetic embryos

作　　者：卢晟盛[1] 魏凤[1] 房慧伶[2] 陆阳清[1] 卢克焕[1] 

机构地区：[1]广西大学广西亚热带生物资源保护利用重点实验室 [2]广西大学动物科技学院,广西南宁530004

出　　处：《福建农林大学学报（自然科学版）》2007年第3期279-283,共5页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家自然科学基金资助项目(30660127);广西科学基金资助项目(0640002);广西亚热带生物资源保护利用重点实验室-省部共建国家重点实验室培育基地开放项目(SB0505)

摘　　要：猪的卵巢卵母细胞在含不同剂量葡萄糖+丙酮酸钠的成熟培养基中体外成熟培养44-48 h,检查核成熟率(试验一),进行化学法孤雌激活后,在含不同浓度的乙二胺四乙酸钠(EDTA-Na)和不同剂量的葡萄糖+谷氨酰胺+牛磺酸+亚牛磺酸的培养基中进行体外培养,检查孤雌激活胚第48小时的卵裂率和第168小时的囊胚发育率(试验二),研究成熟培养基中葡萄糖+丙酮酸钠剂量对卵母细胞体外成熟核成熟率的影响及培养基中不同浓度EDTA-Na和葡萄糖+谷氨酰胺+牛磺酸+亚牛磺酸的剂量对孤雌激活胚胎体外发育的影响.结果表明:(1)葡萄糖+丙酮酸钠在1倍剂量时,猪卵母细胞体外成熟的核成熟率为(59.3±5.0)%;当剂量加倍时,核成熟率极显著下降(P<0.01),为(51.0±4.4)%.(2)当EDTA-Na浓度增高,囊胚发育率上升,当浓度达50μmol.L-1时囊胚发育率最高,为(7.2±4.1)%,此后随浓度升高囊胚发育率下降;添加适当浓度(25-100μmol.L-1)EDTA-Na对猪孤雌激活胚的囊胚发育有利(P<0.05),当浓度达200μmol.L-1时其有利作用消失.(3)培养基中葡萄糖+谷氨酰胺+牛磺酸+亚牛磺酸的剂量过高对卵裂率无显著影响(P>0.05),但对囊胚发育不利(P<0.05).可见:(1)适当浓度(25-100μmol.L-1)EDTA-Na对猪早期胚胎体外发育具有促进作用;(2)成熟培养基中能量基质剂量过高会抑制猪卵母细胞体外成熟;(3)胚胎培养基中葡萄糖+谷氨酰胺+牛磺酸+亚牛磺酸的剂量过高会抑制猪早期胚胎的体外发育.The objectives of this study was （ 1 ） to investigate the effect of different concentrations of glucose and sodium pyruvate in maturation medium on the nuclear maturation rate of porcine oocytes matured in vitro; （2） to investigate the effect of different concentrations of glucose, glutamine, taurine and hypotaurine on the in vitro development of porcine parthenogenetic embryos ; （3） to investigate the effect of different concentrations of ethylenediaminetetraacetic acid （EDTA-Na） on the in vitro development of porcine parthenogenetic embryos. Porcine oocytes from abattoir ovaries were matured in maturation medium containing different concentrations of glucose ＋ sodium pyruvate for 44 - 48 hours and their nuclear maturation rates were recorded （ Expt. 1 ） ; after maturation in vitro and chemically parthenogenetic activation, oocytes were cultured in NCSU-23 medium containing different concentrations of glucose ＋ glntamine ＋ taurine ＋ hypotaurine, and different concentrations of EDTA-Na, and the cleavage and blastocyst rates of oocytes were recorded on day 2 and 7 respectively （ Expt. 2）. The results showed that ： （ 1 ） the nuclear maturation rate of porcine oocytes matured in medium containing twice concentration of glucose ＋ pyruvate was significantly lower than that in medium containing once concentration [ （51.0±4.4）% vs （59.3 ±5.0）%, P 〈0.01] ; （2） as EDTA-Na concentration increased, the blastocyst rate of porcine parthenogenetic embryos increased and the highest blastocyst rate [ （7.2± 4.1 ）% ] was obtained at 50 μmol·L^-1 EDTA-Na over which blastocyst rate decreased; （3） addition of a certain amount （25 - 100μmol·L^-1） of EDTA-Na to culture medium was beneficial to the blastocyst formation of porcine parthenogenetic embryos （ P 〈0.05 ） , over which concentration the beneficial effect disappeared; （4） relatively high concentration of glucose ＋ amino acids in culture medium had no effect on the cleavage rate of porcine parthe

关 键 词：乙二胺四乙酸钠(EDTA-Na) 能量基质 氨基酸 猪 卵母细胞 孤雌激活 胚胎 

分 类 号：Q954.4[生物学—动物学]