乙二胺四乙酸钠、能量基质、氨基酸对猪卵母细胞的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] 兰宗宝[1] 房慧伶[2] 陆阳清[1] 卢克焕[1] 王爱德[2] 

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,广西南宁530005 [2]广西大学动物科学技术学院,广西南宁530005

出　　处：《广西农业生物科学》2006年第B09期188-189,共2页Journal of Guangxi Agricultural and Biological Science

基　　金：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课题(SB0505);广西科学基金项目(桂科青0640002和桂科青0447004);广西科学研究与技术开发计划项目(桂科攻0537022-01和桂科攻0330004-14);广州市华粤行仪器有限公司南宁办事处计划内横向项目

摘　　要：本研究探讨了：（1）成熟培养基中葡萄糖和丙酮酸钠剂量对猪卵母细胞体外成熟极体排放率的影响；（2）培养基中葡萄糖、谷氨酰胺、牛磺酸和亚牛磺酸的剂量对猪孤雌激活胚胎体外发育的影响，同时探讨不同浓度乙二胺四乙酸钠（EDTA—Na）对猪孤雌激活胚胎体外发育的影响。利用屠宰场猪卵巢卵母细胞，在含不同剂量葡萄糖十丙酮酸钠的成熟液中体外成熟44～48h，计算其极体排放率（试验1），进行化学法孤雌激活后，在含不同剂量的葡萄糖＋谷氨酰胺＋牛磺酸＋亚牛磺酸与不同浓度的EDTA—Na的培养基中体外培养，计算孤雌激活胚的第48小时卵裂率和第168小时囊胚率（试验2）。结果表明：（1）葡萄糖＋丙酮酸钠在l倍剂量（浓度分别为3．05mmol／L和0．91mmol／L）时，猪卵母细胞体外成熟的核成熟率为（59．3±5．0）％，当其剂量加倍时，猪卵母细胞体外成熟的核成熟率极显著下降（P〈0．01），为（51．0±4．4）％；（2）随着EDTA—Na添加浓度的增加，猪孤雌激活胚的的囊胚发育率不断上升，添加浓度达到50μmol／L时囊胚率最高（7．2±4．1）9／6，超过此浓度后囊胚率开始下降，培养基中添加适当浓度（25—100μmol／L）的EDTA—Na对猪孤雌激活胚的囊胚发育率具有有利作用（P〈0．05），添加浓度达到200μmol／L时其有利作用消失；（3）培养基中葡萄糖＋氨基酸的剂量过高（葡萄糖、谷氨酰胺、牛磺酸、亚牛磺酸的浓度分别大于5．55，1．0，7．0，5．0mmol／L）对猪孤雌激活胚的卵裂率无显著影响（P〉0．05），但对其囊胚发育率有不利作用（P〈O．05）。本研究得出以下结论：（1）适当浓度（25～100μmol／L）的EDTA—Na对猪早期胚胎体外发育具有促进作用；（2）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）培养基中能量基质十The objectives of this study was to investigate： （1） The effectof different concentrations of glucose and sodium pyruvate in maturation medium on the nuclear maturation rate of porcine oocytes matured in vitro; （2） The effects of different concentrations of glucose, glutamine, taurine and hypotaurine on the in vitro development of porcine parthenogenetic embryos; （3） The effects of different concentrations of ethylenediaminetetraacetic acid （EDTA） 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＋glutamine＋taurine＋hypotaurine, and different concentrations of EDTA- Na, and the cleavage and blastocyst rates of oocytes were recorded after 48 h and 168 h respectively （Expt. 2）. The results showed that： （1） The nuclear maturation rate of porcine oocytes matured in the medium containing twice concentration of glucose＋pyruvate was significantly lower than that in the medium containing once concentration of which the concentration of glucose and pyruvate were 3.05 and 0.91 mmol/L respectively [ （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 50μmol/L EDTA-Na over which blastocyst rate decreased; （3） Addition of a certain amount （25-100μmol/L） 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 （the concentrations of glucose,

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

分 类 号：Q813[生物学—生物工程]