哺乳动物超数排卵和卵母细胞的生发泡互换分析-analysis of the exchange between superovulation and oocyte foaming in mammals.docxVIP

摘要一、家猫的超数排卵及卵母细胞的体外成熟对家猫超数排卵(superovulation)、卵母细胞的体外成熟(in vitro maturation。IVM)、孤雌激活(parthenogenetls)进行了研究，结果表明：FSH／hCG 法平均超排卵数(9．15枚／只)，显著多于PMSG／hCG(5．28枚／只)法(Po．01)，阴 道刺激对猫超排有显著促进作用；TCM．199+10％FCs+PMSG对家猫卵巢卵 母细胞进行体外成熟培养，其成熟率为78．9％(116／147)，显著高于在 TCM．199+10％FCS 中的成熟率48．8％(66／135) (P0．01)； 采用 0．5kV／cm，1．0kv／cm和1．5kV／cm三种直流电压以相同的脉冲时间40ps，脉冲次数3次对猫卵进行孤雌激活，1．5kV／cm的电压对猫卵激活率(88．7％)显著高于1．0kV／cm(62．2％)和0．5kV／em(45．4％)的激活率(PO．01)．二、兔初级卵母细胞生发泡移植方法的研究本文共收集GV期卵母细胞987枚，GV互换成功率27．6％(272／987)，融合 率为84．6％(230／272)，重构卵的成熟率(排出第一极体)77．4％(178／230)。 经ICSI后，受精率为70．8％(126／178)。这些结果说明，显微操作和电融合过 程并没有影响移植后的GV卵的成熟发育，也不影响作为受体的卵胞质的形态 结构， 从而证明对兔初级卵母细胞进行生发泡互换是可行的。三、兔生发泡移植的重构卵经显微受精后产仔本文以兔为实验动物模型进行GV互换。当进行同卵或异卵Gv移植时， 有约80％的重构卵成熟至MII期。将这些成熟的重构卵进行显微受精(ICSI) 时，有7．1-9．4％的胚胎发育到囊胚。将93枚受精卵移植到6只受体母兔后，获 得2只仔兔，这是世界上首批通过GV互换、体外成熟和ICSI后，经胚胎移植 获得的动物。关键词：家猫，超数排卵，体外成熟，孤雌激活，家兔，生发泡， 生发泡移植，胞质内精子注射Abstract1．Superovulation And砌Vitro Maturation of Domastic CatSuperovulation，in vitro maturation and parthenogenetic activation of domestic cat oocytes were studied。The results showed that an average 9．1 5 eggs were ovulated by the method of FSH／hCG administration， evidently higher than that obtained by injecting PMSG／hCG f5．28)． Stimulation of vagina promoted the superovultion significantly(P0．01)． The maturation rate in vitro of domestic cat ovary oocytes in TCM-199+10％FCS+PMSG was 78．9％． significently higher than that in TCM-199(48．8％)+10％FCS(Po．01)；the activation rate of cat oocytes was 88。7％by using 1．5kWcm，409s，3 times electric stimulation， significently higher than that obtained by using 1．0kWcm，409s，3 times(62．2％)and 1．0kV／cm，409s，3 times(45．4％)(Po．01)。I2．Technique For Germinal Vesicle Transfer From RabbitPrimary OoeytesIn order toevaluate the feasibility of the germinal vesiclerGV、transfer using rabbit as a model．Wb collected total 987 GV stakeoocytes from the ovaries ofⅥmite Japan Big Ear rabbits 72h after FSH injection．Exchanged the GV-karyoplast between the primary oocytes， fused by electrical pulses，and then m