试验探讨了水牛和黄牛同种及种间显微授精的可行性。体外成熟22~24h的水牛和黄牛卵母细胞分别注入冷冻/解冻的尼里-拉菲水牛和利木赞黄牛精子,进行同种或种间的显微授精操作,构建的ICSI胚胎一部分培养到16~18h固定染色检查原核形成情况,另外一部分胚胎培养2~9d分别记录胚胎的分裂情况及囊胚发育情况。结果发现,水牛同种(水牛+水牛)和水牛异种(水牛+黄牛)显微授精胚胎的双原核率(47.73%和36.67%)、分裂率(68.00%和72.64%)和囊胚发育率(16.44%和22.39%)均无显著差异(P〉0.05);黄牛同种(黄牛+黄牛)和黄牛异种(黄牛+水牛)显微授精胚胎的双原核率(60.00%和45.28%)、分裂率(73.33%和71.31%)和囊胚发育率(28.57%和33.70%)亦无显著差异(P〉0.05)。以上结果表明:①水牛精子注射到黄牛卵子和黄牛精子注射到水牛卵子的种间授精胚胎均可以体外发育到囊胚阶段;②黄牛卵子无论同种和种间显微授精的效果均优于水牛卵子。
The possibility of intraspecific and interspecific fertilization between buffalo and cattle was explored in this study.Buffalo and bovine oocytes matured in vitro for 22 to 24 h were fertilized by injection of buffalo or bovine frozen-thawed spermatozoa.At 16 to 18 h after injection,a portion of ICSI oocytes were fixed in acetic ethanol to examine the pronuclear formation,others were cultured in vitro to evaluate their cleavage rate and embryonic developmental ability.There was no difference in the two pronuclear formation rate(47.73% and 36.67%),cleavage rate(68.00% and 72.64%) and blastocyst development(16.44% and 22.39%) of buffalo oocytes fertilized with buffalo(intraspecies) and bovine sperm(interspecies)(P0.05).The two pronuclear formation rate(60.00%),cleavage rate(73.33%) and blastocyst rate(28.57%) of bovine oocytes fertilized with bovine sperm(intraspecies) were also not different with the embryos from those(45.28%,71.31% and 33.70%,P0.05) of bovine oocytes fertilized with buffalo sperm(interspecies).This results indicated that: ① Buffalo oocytes fertilized with bovine sperm and bovine oocytes fertilized with buffalo sperm could develop to blastocysts,②The developmental capacity of bovine oocytes was higher than buffalo oocytes as evidenced by their proportion cleaved and developed to blastocysts after intraspecific and interspecific fertilization by ICSI.