为探讨昆明白小鼠不同类型的卵胞质对小鼠体细胞核移植胚胎发育的影响，采用C57BL/6小鼠耳成纤维细胞为核供体，昆明白小鼠卵母细胞为核受体进行单次核移植，同时采用将重构胚类合子核移入去核合子、重构胚2-细胞卵裂球细胞核移入去核MⅡ期卵的方法进行连续核移植，通过将重构胚2-细胞卵裂球与正常昆明白小鼠2-细胞胚胎卵裂球交换融合得到四倍体胚胎对所得胚胎进行体外培养，结果发现，以去核的昆明白小鼠MⅡ期卵母细胞为核受体进行单次核移植所得到的重构胚胎发育率很低，且只能发育到8-细胞阶段两种连续核移植重构胚发育率均有所提高，但仅重构胚类合子核移入去核合子时有囊胚出现，囊胚发育率为1.9%重构四倍体胚胎囊胚发育率却很高（62.3%），染色体分析表明其中51.5%为真正的四倍体结果表明，昆明白小鼠不同类型核受体对重构胚胎的发育能力有显著影响，其去核卵母细胞胞质对体细胞核重编程的能力较弱，而当其与正常受精卵胞质共同作用时可较好地对体细胞核重编程。

In order to study effects of electro-fusion and strontium chloride(SrCl2)activation in nuclear transfer experiment on activation and development of mouse oocytes, concentration and treatment duration of SrCl2, electro-pulse and electro-pulse combining SrCl2 were used to activate mouse oocytes which were obtained after hCG 17h.Activated oocytes were cultured in vitro in CZB medium.The results were as follows: 82.4%activation percentage was obtained when the oocytes were treated with lOmmol/L SrCl2 for 6h, it was significantly(P>0.05)higher than those obtained from that treated with the Smmol/L or lOmmol/L SrCl2 for 4h.The activation percentage was not significantly different between Smmol/L and lOmmol/L SrC12 for 6h, but the percentage of morula and blastocyst in lOmmol/L SrCl2 6h group was significantly(P>O.05)higher than those in Smmol/L SrCl2 6h group.In the groups of treatment with electro-pulse, the best activation percentage(70.9%)was obtained when the oocytes were treated with l.Okv/cm, 320

为探讨体细胞来源及培养代数对核移植重构胚发育的影响，实验采用电融合法将小鼠2-细胞胚胎卵裂球、胚胎干细胞（ES）、胎儿成纤维细胞、耳成纤维细胞、尾尖成纤维细胞、睾丸支持细胞和精原细胞以及不同培养代次的胎儿成纤维细胞进行了核移植。结果显示：2-细胞胚胎卵裂球供核重构胚发育最好，囊胚率为7.4%；ES细胞重构胚虽然发育率低，但仍有囊胚出现，比例为0.7%；胎儿成纤维细胞重构胚最高发育阶段为桑椹胚，比例为0.2%；精原细胞重构胚只能发育到8-细胞阶段，比例为0.3%；其他几类细胞重构胚则仅能发育至4-细胞阶段。不同培养代数的胎儿成纤维细胞重构胚除第3代外都可发育到8-细胞阶段，且发育率差异不显著，但第一代细胞重构胚2-细胞发育率（40.7%）显著低于2、3和4代细胞重构胚。结果表明:不同分化程度的细胞核移植后，重新编程的难易程度是不一样的，分化程度越高则重新编程越难；未调整细胞周期的ES细胞由于多数处于S期，所以重构胚发育率很低；体外培养传代有利于体细胞核移植后重新编程。

绵羊卵巢采自乌鲁木齐市屠宰场，卵母细胞体外成熟24-25h后，由体外获能的附睾精子授精，50h后，将卵裂至#2-8细胞的绵羊胚胎移入添加了不同浓度细胞因子的培养液中继续培养，结果表明，加入5、10、15、µg/ml胰岛素（Insulin）分别得到17.46%、23.81%、22.95%的囊胚率，与对照组9.52%相比差异显著（P<0.05）。高浓度（500µg/ml）的粒细胞巨噬细胞集落刺激因子（Granulocyle-macrophagecolonystimulatingfactor，GM-CSF）可获得21.82%的囊胚率，与对照组7.27%相比差异显著（P<0.05）。胚胎培养液中分别加入10、100、500ng/ml的白细胞介素-3（Interleukin-3，IL-3）、白细胞介素-8（IL-8）和神经生长因子（Nucleargoowthfactor，NGF）与对照组8、NGF相比，囊胚率差异不显著。本研究表明，胰岛素、GM-CSF对绵羊体外受精胚胎早期发育有促进作用，IL-3、IL-8、NGF对绵羊胚胎的早期发育无明显作用。

本研究应用激光扫描共聚焦显微镜的光漂白恢复技术（FRAP）分析兔早期胚胎卵裂球之间通过间隙连接介导的细胞通讯（GJIC）。研究结果发现，用强激光分别将4-细胞期胚胎、异裂胚胎和8-细胞期胚胎的一个卵裂球荧光光漂白后，经过15分钟的荧光恢复，4-细胞期胚胎的光漂白恢复率为17.80%，异裂胚胎的光漂白恢复率为23.7%，二者之间没有明显的差异；8-细胞期胚胎的光漂白恢复率为78.2%，与前二者之间存在明显的差异。推测兔早期胚胎卵裂球细胞间隙连接建立的时间在8-细胞阶段，胚胎卵裂球间隙连接通讯可能是兔胚胎正常发育的重要条件。

Adult somatic cell nuclear transfer was conducted by using cultured ear fibroblast cells obtained from a Holstein female cow(GN)and a Galoway herd bull(GLV).The percentages of reconstructed eggs developed into blastocysts were similar in GN(23.98%, 123 of 513)and in GLV groups(29.55%, 138 of 467).However, the rate of reconstructed female(GN)embryos developed into term was higher than that of male(GLV)(8.02%and 1.82%, respectively).Three kinds of cows, Luxi Yellow cows, Holstein heifers and Holstein cows with normal reproductive records were used as recipients.When the reconstructed embryos from GN were transferred, there was no difference in the pregnancy rate among three kinds of recipients, but the abortion rate of Luxi Yellow cows was significantly higher(85.71%)than in the other two groups(14.29%and 0%, respectively; P<0.05).And the percentages of newborn calves in transferred embryos were significantly different between Luxi Yellow cows and Holstein breed(1.54%, 10.39%and 20.0%, respectively, P<0.05). However, when reconstructed embryos from GLV were transferred, there was no difference among three kinds of recipients in the pregnancy rate, the abortion rate and the delivery rate.

利用体外培养的成年母牛（GN）和公牛（GLV）的外耳成纤维细胞作为核供体，进行了规模较大的克隆牛研究，并成功地获得了一批成年体细胞克隆牛犊。经核移植构建的重构胚的囊胚发育率分别为23.98%（GN，123/513）和29.55%（GLV，138/467）。然而，雌性重构胚（GN）发育满期的比例却明显高于雄性重构胚（GLV），分别为8.02%和1.82%。比较了鲁西黄牛、青年Holstein奶牛及经产Holstein奶牛种3不同受体牛的移植效果.在移植GN重构胚时，3种受体牛的妊娠率无显著差异，但黄牛的流产率高达85.71%，显著高于青年奶牛及经产奶牛的流产率（分别为14.29%和0%，P<0.05）。黄牛受体生产的克隆牛犊占移植胚胎总数的1.54%，显著低于青年Holstein奶牛及经产Holstein奶牛（分别是10.39%及20.0%，P<0.05）。而移植GLV重构胚时，3种受体牛的妊娠率、流产率及产犊比率均无显著差异

The completion of meiosis requires the spatial and temporal coordination of cytokinesis and karyokinesis. During meiotic maturation,many events,such as formation, location, and rotation of the meiotic spindle as well as chromosomal movement,polar body extrusion, and pronuclear migration, are dependent on regulation of the cytoskeleton system.To study functions of microfilaments in meiosis, we induced metaphase II(MII) mouse oocytes to resume meiosis by in vitro fertilization or parthenogenetic activation, and we treated such oocytes with cytochalasin B (CB). The changes of the meiotic spindle,as visualized in preparations stained for β-tubulin and chromatin,were observed by fluorescent confocal microscopy. The meiotic spindle of MII oocytes was observed to be parallel to the plasmalemma.After meiosis had resumed,the spindle rotated to the vertical position so that the second polar body could be extruded into the perivitelline space.When meiosis resumed and oocytes were treated with 10μg/ml of CB, the spindle rotation was inhibited.Consequently, the oocyte formed an extra pronucleus instead of extruding a second polar body. These results indicate that spindle rotation is essential for polar body extrusion; it is the microfilaments that play a crucial role in regulating rotation of the meiotic spindle.