作　　者： ; ; (石俊松）; (罗绿花）; (周荣）; (周秀）; (麦然标）;

机构地区： 广东温氏食品集团有限公司研究院广东新兴527439

出　　处： 《华南农业大学学报》 2012年第3期388-392,共5页

摘　　要： 从猪卵巢上获取的卵母细胞经体外培养成熟42～44 h后去除颗粒细胞,选取成熟的卵母细胞进行孤雌激活培养.试验采用不同的场强、脉冲时程和脉冲次数进行对照试验,并比较了细胞松弛素B(CB)辅助激活对孤雌胚胎发育的影响.结果显示:1)当场强为0.8 kV.cm-1,选取4个脉冲时程(80、100、120、140μs)参数分别进行1次、2次脉冲激活时,100μs、1次脉冲囊胚率最高,为(42.22±5.38)%,但同其他组无显著差异(P>0.05).2)脉冲时程为100μs,选取4个场强(0.8、1.0、1.2、1.4 kV.cm-1)参数分别进行1次、2次脉冲激活时,1.4 kV.cm-1、1次脉冲囊胚率最高,为(44.04±0.68)%,但同其他组无显著差异(P>0.05).3)场强为0.8 kV.cm-1和1.4 kV.cm-1,选取4个脉冲时程(80、100、120、140μs)进行1次脉冲激活时,1.4 kV.cm-1+80μs组囊胚率(54.60±2.86)%均高于其他组,且极显著高于1.4 kV.cm-1+140μs、0.8 kV.cm-1+80μs和0.8 kV.cm-1+140μs组(P<0.01).4)用1.4 kV.cm-1、80μs、1次脉冲激活卵母细胞后,使用CB辅助激活4 h的囊胚率为(54.07±3.12)%,极显著(P<0.01)高于不使用CB组.以上结果说明,电激活中脉冲次数对猪孤雌胚胎发育无显著影响,且在本试验条件下,采用1.4 kV.cm-1的场强、80μs的脉冲时程,并使用CB辅助激活4 h,能够得到较高囊胚率的猪孤雌激活效率. Oocytes were collected from porcine ovaries, cultured in vitro for 42 - 44 h, and cumulus cells were removed from oocytes. Matured oocytes were selected for parthenogenetical activation. The effect of different field strengths （FSs）, pulse durations （PDs）, electrical pulse numbers （EPNs）, and the influence of cytochalasin B treatment on development of parthenogenetically activated porcine oocytes were investigated. In the first experiment, 4 different PDs （80,100,120,140 μs） were applied with a single or two 0. 8 kV ·cm-1 pulse（s） to activate the oocytes. The results showed that a single 100 μs pulse of 0. 8 kV/cm resulted in the highest blastocyst rate of （42.22 ± 5.38 ） %, but it was not statistically different from the blastocyst rates of other groups （P 〉 0. 05）. In the second experiment, 4 different FSs （0. 8, 1.0,1.2,1.4 kV·cm-11 ） were used with a single or two 100 μs pulse（s） to stimulate the oocytes. The highest blastocyst rate [ （44.04 ±0. 68）% 1 was observed in the group applied with single 100 μs pulse of 1.4 kV ·cm-1, but it was not statistically different from the blastoeyst rates of other groups （ P 〉 0. 05）. In the third experiment, 4 different PDs （80,100,120,140 μs） were used with a single 0. 8 kV·cm-1 or 1.4 kV·cm-1 pulses to activate the ooeytes. The blastoeyst rate of the group treated with a single 80 μs pulse of 1.4 kV ·cm-1 reached （54.60 ± 2. 86） % , which was significantly higher than that of other groups （P 〈 0. 01 ）. In the fourth experiment, one group of oocytes was stimulated only by a single 80 μs pulses of 1.4 kV·cm-1 , while the other groups was activated with a single 80 μs pulses of 1.4 kV ·cm-1 followed by 4 h-cytoehalasin B treatment. The results indicated that the blastoeyst rate of the latter group was （54. 07 ± 3.12）%, significantly higher than that of the former group （P 〈 0. 01 ）. The above results suggested that EPN had no significant effect on parthenogenetical development of por

关 键 词： 猪卵母细胞 囊胚率 孤雌激活

领　　域： [农业科学] [农业科学]