不同激活剂对兔卵母细胞孤雌激活效果的研究-论文.pdf

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1、江西农业学报2014，26(9)：90—93ActaAgricuhuraeJiangxi不同激活剂对兔卵母细胞孤雌激活效果的研究王鸽，张君涛，张胜军，魏莉莉，曹书郡，徐秋勤，柳文龙，刘童，张志平一(河南农业大学牧医工程学院，河南郑州450002)摘要：目的：探寻一种稳定有效的卵母细胞激活方法，为兔体细胞核移植提供实验基础。方法：兔超排后获得卵母细胞体外成熟22—24h，经脱颗粒处理后采用不同激活剂和不同处理时间进行激活，体外培养5—7d，观察胚胎发育与囊胚形成情况。结果：卵母细胞经过离子霉素作用5min或者乙醇作用7min均能被激活，但两者卵裂率、囊胚率都较低；乙醇和离子霉素分别与6一D

2、MAP联合激活兔卵母细胞时，离子霉素+6一DMAP处理组卵裂率(73．7％)显著高于乙醇+6一DMAP处理组(38．5％，P<0．05)；与离子霉素联合激活时，2．0mmol／L的6一DMAP处理5．0h时的卵裂率和囊胚率最高，达75．6％和45．5％。结论：激活方法对兔卵母细胞孤雌激活率的影响很大，不同激活剂的联合应用及激活剂的作用时间对兔卵母细胞的激活存在显著差异。关键词：兔卵母细胞；孤雌激活；离子霉素；乙醇；6一二甲基氨基嘌呤中图分类号：$829．1文献标志码：A文章编号：1001—8581(2014)09—0090—04StudyonEfectofDifferentStimuli

3、onParthenogeneticActivationofRabbitOocytesWANGGe，ZHANGJun—tao，ZHANGSheng～jun，WEILi—li，CAOShu—jun，XUQiu—qin，LIUWen—long，LIUTong，ZHANGZhi—ping(CollegeofAnimalandVeterinaryScience，HenanAgriculturalUniversity，Zhengzhou450002，China)Abstract：Theobiectiveofthisstudyistoseekastableandeffectiveme山odforthe

4、parthenogeneticactivationofrabbitoo．cytes．andmakeanexperimentalfoundationforthesomaticcellnucleartransferofrabbit．Cumulus—oocytecomplexeswereharves．tedfromtheovariesofsuperovulatedrabbits，andwerematuredinvitrofor22～24h；afterremovingthecumuluscells。theoocyteswereactivatedbyvariousstimulifordiffere

5、nttime．Afterinvitroculturefor5—7d．theparthenogeneticdevelopmentofstimulatedoocyteswasassessedbymicroscopicobservation．Theresultsshowedthatrabbitoocytescouldbeactivatednotonlybyionomycintreatmentfor5min，butalsobyethanoltreatmentfor7min，buttheircleavagerateandblastularateallwererelativelylow．When6一

6、DMAPcombinedwithionomycinorethanolwasusedtoactivaterabbitOOCytes．thecleavagerate(73．7％)inthetreatmentofionomycin+6一DMAPwassignificantly(P<0．05)higherthanthat(38．5％)inthetreatmentofethanol+6一DMAP；thetreatmentofionomycin+2．0mmol／L6一DMAPfor5．0hobtainedthehighestcleavagerateandblastularate．being75．6％

7、and45．5％respectively．Itwasconcludedthatactivationmethodhadgreatinfluenceontheparthenogeneticactivationrateofrabbitoo．cytes，andthereweresignificantdifferencesinactivationeffeatamongdifferenttreatments(combinedutilizatio