猪卵母细胞与猪、水牛及食蟹猴精子的胞质内显微受精

Intracytoplasmic injection of porcine,buffalo and cynmolgus monkey spermatozoon into porcine oocytes

为探讨猪精子、水牛精子及食蟹猴精子分别注入到猪卵母细胞后原核形成及早期胚胎发育情况,利用屠宰场收集的猪卵母细胞,经体外成熟44～48h后,进行胞质内显微受精(ICSI)操作。试验1:体外培养18～19h,用Ho-echst33342荧光染色,检查原核形成情况。试验2:进行体外培养,ICSI后2d检查分裂率,7d记录囊胚率。试验1结果显示:在原核形成率上,猪同种显微受精,原核形成率(50.10%)显著高于注入水牛精子(37.06%)及食蟹猴精子(37.48%),且差异显著(P<0.05),注入水牛精子和食蟹猴精子间差异不显著。试验2结果显示:猪同种显微受精所得的分裂率(86.58%)和囊胚率(29.93%)与水牛精子注入(70.98%,18.48%)、食蟹猴精子注入(78.69%,16.92%)差异显著(P<0.05)。结果表明,水牛精子、食蟹猴精子分别注入到猪卵母细胞后,观察到雌雄原核和精子解聚;水牛精子注入猪卵母细胞与食蟹猴精子注入猪卵母细胞,经体外培养发育到囊胚。

To determine the incidence of pronuclear formation and in vitro development of porcine oocytes following intracytoplasmie injection using the sperms of pig,cynomolgus monkey and buffalo,the porcine oocytes from abattoir ovaries were matured in vitro for 44-48 h,and ICSI （intracytoplasmin sperm injection） were activated and then cultured for 18-19 h for staining by Hoechst 33342 to assess pronuelear formation （Expt 1）. The cleavage and blastocyst rates were examined on day 2 and day 7 respectively （Expt 2）. The results showed that：in the experiment 1 ,the rate of pronuclear formation of injecting porcine spermatozoa group （50.10 % ） was significantly higher than that of injecting cynomolgus monkey （37. 48 %） and buffalo spermatozoa （37. 06%） groups. There was no significant difference on it between the two foreign species spermatozoa. In the experiment 2, there was significant difference （P 〈0.05） among injecting porcine （86.58% ;29.93%） ,cynomolgus monkey （78. 690% ;16.92%） ,buffalo （70.98% ; 18.48%） sperm groups on cleavage and blastocyt rates. In conclusion,the pronuclear and blastocyt formation were observed in the porcine oocytes injected with spermatozoa of eynomolgus monkey and buffalo.