摘要
以成年家兔上皮细胞为核供体,对兔体细胞核移植中孤雌激活、去核、融合、激活及重构胚发育等环节中的有关参数进行筛选和研究.证实160、200和240 V/mm电场强度时的孤雌激活率(77.78%、95.12%、97.30%)无显著差异;200和240 V/mm电场强度时的孤雌激活胚囊胚率(52.94%、48.64%)显著高于160 V/mm时(16.67%);200和240 V/mm电场强度时的重构胚融合率(82.85%、79.78%)无显著差异;200 V/mm电场强度时的重构胚分裂率(71.69%)显著高于240 V/mm时(56.56%),裂解率(6.82%)显著低于240 V/mm时(17.92%).在纺锤体观测仪下,第一极体与纺锤体在一起或相距较近的占60%左右.原核明显的重构胚的分裂率(91.75%)极显著的高于观察不到原核的重构胚(42.86%).重构胚体外培养发育囊胚率(24.39%)低于受精卵原核胚的体外培养的发育率(86.05%),但两者同自然交配体内发育的囊胚的细胞数(81±17)差异不显著.将646枚2~4细胞重构胚移植到38只同期化或非同期化的受体后没有幼仔出生.
This paper screened and optimized some parameters of nuclear transfer on rabbits, in cluding parthenogenesis, fusion, activation, and development of NT embryos, the nuclear donors were skin epidermal cells of adult rabbits. The parthenogenesis rates were no significant difference, under the electric field of 160 V/mm (77.78%), 200 V/mm (95.12%), and 240 V/mm (97.30%)1 the blastocyst rates under 200 V/mm (52.94%) and 240 V/mm (48.64%) were significantly higher than that under 160 V/mm (16.67%);the fusion rates of NT embryos were no significant difference under 200 V/mm (82.85%) and 240 V/mm (79.78%); the cleavage rate under 200 V/mm (71.69%) was significantly higher than that under 240 V/mm (56.56%), the collapse rate under 200 V/mm (6. 82%) was significantly lower than that under 240 V/ram (17.92%). There was about 60% that the first polar body was together with and close to spindle under spindleview imaging system. The cleavage rate with visible pro nuclears zone (91. 75%) was very significantly higher than that without visible pro-nuclears zone (42.86%). The blastocyst rate (24.39%) of NT embryos was lower than developmental rate (86.05%) of embryos of pro-nuclear zygote in vitro, but both blastocyst cells were no significant difference with those (81±17) of spontaneous mating and development in vivo. Implanted 646 2-4-cell NT embryos into 38 synchronized or asynchronized recipients, no fetus was born.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2006年第5期441-446,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(30070374)
"863"计划基金(2002AA216161)
国家重点基础研究发展计划基金(G199954301)
关键词
兔
上皮细胞
卵母细胞
孤雌激活
核移植
rabbits
epidermal cells
oocytes
parthenogenesis
nuclear transfer