作　　者： ; ;

机构地区： 江苏省农业科学院

出　　处： 《畜牧兽医学报》 1994年第6期507-512,共6页

摘　　要： 本文分析了影响兔卵母细胞电激活及发育的主要因素，建立了激活及体外发育的实验程序。试验表明，激活率随着卵龄的增加而升高，采用０．３Ｍ甘露醇为电激液时２０ｈ卵龄的激活率为４０％，显著高于１６及１８ｈ的１４％（Ｐ＜０．０１）和２０％（Ｐ＜０．０５）。会电解质成分的电激液的激活率显著高于非电解质液。随着电激次数及场强的增加，激活率亦显著升高，以０．３Ｍ甘露醇，１００μＭＣａＣｌ＿２＋１００μＭＭｇＣｌ＿２，施加１．６５ｋｖ／ｃｍ，８０μｓ，间隔３０ｍｉｎ进行３次和６次电刺激时的激活率分别为７９％和８３％，显著高于１次电激时的５３％，Ｐ＜０．０１，而当电压升高至２．１０ｋｖ／ｃｍ时，６次电激后的激活率则为１００％，Ｐ＜０．０５。激活胚小：７Ｕ．７～８４．５％形成具有２个原核及１个极体的二倍体。激活胚在最佳电脉冲参数条件下，以改良ＴＣＭ１９９＋２５ｍＭＨＥＰＥＳ＋１０％ＦＣＳ为操作液时的囊胚发育率为２６．６％，当以改良ＴＣＭＩ９９＋ＮａＨＣＯ３２．２ｇ／Ｉ＋１０％ＦＣＳ为操作液时的囊胚发育率则为５７．１％，囊胚孵化率为４６．４％，Ｐ＜０．０１。 In this paper, the factors affecting parthenogenetic activation by electrical pulsingand development in vitro were analyzed ,and the experimental procedure for activationand development in vitro were established.The results were as follows: In comparingoocyte age,20-h post HCG treatment oocytes activated at a higher rate(40%)than l6-hand l8-h post HCG oocytes(14%P<0.0l,and 20%P<0.05),when 0.3M mannitollvas used as pulsing medium. The activation rate of oocyte pulsed in eletrolyte mediumis significantly higher than that in nonelectrolyte medium,P<0.05.A higher proportionof oocvtes activated as pulse number and field strength increased.When the oocytesvere pulsed in the medium of 0.3 M mannitol + 100pM CaCl2+100μM MgCl2 with thepulsing parameter:duration 80μs,field strength l.65kv/cm,the activatiOn rates of 3pulses and 6 pulses at 30 minute intervals(79%, 83%)were higher than that of lpulse(53%,P<0.01).A dramatic increase of activation rate was obtained at 2.10kv/cm,6 pulses in 30 minute intervals(100%,P<0.05).76.7～87.5%of the activationoocvtes were diploid showing two pronuclear and one polar body.The effect of manipulation medium on in vitro development ability of activated oocytes was investigated.26.6%of the activated oocytes developed to blastocyst stage in the optimal parametersof pulses in modified TCMI99+25mM HEPES+10%FCS,hOWever,57.l%and 46.4%of the activated oocytes developed to blastocyst stage and hatching blastocyst stagerespectively in modified TCMI99+NaHCO,2.2g/1 + 10%FCS,P<0.0l.

关 键 词： 兔 卵母细胞 电刺激 孤雌生殖 激活

领　　域： [农业科学] [农业科学]