畜牧兽医学报 ›› 2011, Vol. 42 ›› Issue (2): 182-189.doi:

• 遗传繁育 • 上一篇    下一篇

羔羊卵泡诱导发育、活体采卵和体外发育潜能研究

胡鹏飞1,2,张贵学1*,于国庆2,吴增华2,李向臣3,关伟军3*   

  1. 1. 东北农业大学动物科学技术学院,哈尔滨 150030;2. 辽宁省农业科学院风沙地改良利用研究所,阜新 123000;3. 中国农业科学院北京畜牧兽医研究所,北京 100193
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-02-25 发布日期:2011-02-25
  • 通讯作者: 张贵学;关伟军

Study on Induction, in vivo Pick-up and in vitro Development of PrepubertalLambs Oocytes

HU Peng-fei1,2, ZHANG Gui-xue1*, YU Guo-qing2, WU Zeng-hua2, LI Xiang-chen3, GUAN Wei-jun3*   

  1. 1.College of Animal Science and Technology, Northeast Agricultural University, Harbin150030, China;2.Institute of Windy Sands Improvement Technology, Liaoning Academy ofAgricultural Sciences,Fuxin 123000, China; 3. Institute of Animal Science,Chinese Academy of Agricultural Sciences, Beijing 100193, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-02-25 Published:2011-02-25

摘要: 本研究旨在探讨聚乙烯吡咯烷酮(Polyvinyl pyrrolidone, PVP)作为缓释剂在羔羊卵泡诱导发育中的应用效果,以及品种、年龄、激素剂量和超排次数对活体采集卵母细胞数量和体外发育能力的影响。选用4~12周龄无角道塞特、萨福克母羔120只,分为15%PVP一次肌注组、30%PVP一次肌注组和常规递减注射组,每组40只,注射剂量均为120 IU,对30%PVP一次肌注组按品种、周龄、激素剂量和重复次数分为3~4小组,分别进行卵泡诱导发育和活体采卵,并与屠宰厂采集的卵母细胞对比进行体外受精和胚胎移植。结果表明,采用30% PVP缓释FSH进行羔羊超排,可以起到与常规超排一致的效果,二者差异不显著(P>0.05),但采用15% PVP缓释FSH进行羔羊超排没有取得理想的效果,在各项指标上均显著低于30% PVP组和常规递减注射组(P<0.05);采用30% PVP缓释FSH分别超排道塞特和萨福克羔羊,只均获卵数和可用卵数差异不显著(P>0.05),4、6和8周龄羔羊只均获卵数和可用卵数显著高于12周龄羔羊组(P<0.05),40 IU组只均获卵数显著低于80、120和240 IU剂量组(P<0.05),240 IU剂量组只均获卵数最高,但与120 IU剂量组相比差异不显著(P>0.05),重复超排后只均获卵数和可用卵数显著低于初次超排(P<0.05),同时超排成功率也显著下降(P<0.05);与屠宰场采集的卵母细胞相比,活体采集的羔羊卵母细胞体外成熟率、卵裂率、桑椹胚率和囊胚率均较低(P<0.05),但受精率二者差异不显著(P>0.05);分别冷冻保存了95和140个羔羊体外受精胚胎与屠宰场体外受精胚胎,复苏后存活率分别为95.8%和94.3%,随后进行了胚胎移植,移植后的妊娠率二者差异不显著(P>0.05),分别产羔2和7只。结果提示,采用30%PVP缓释技术进行羔羊卵泡诱导发育,可以取代常规的递减注射法;年龄、激素剂量和超排次数影响活体采集的卵母细胞数量,4周龄的道塞特羔羊采用30%PVP缓释120 IU FSH可得到最高的只均获卵数和可用卵数,活体采集的羔羊卵母细胞可经体外受精和胚胎移植获得后代,具有与屠宰场采集的成羊卵母细胞相同的体外发育能力。

Abstract: This experiment was conducted to study the effect of Polyvinyl pyrrolidone (PVP) on superovulation of prepubertal lambs, and the effects of factors such as breed, age, hormone dose and frequency of ovum pick-up on number of oocytes obtained were also studied. One hundred and twenty 4.12 weekold Poll Dorset and Suffolk lambs were assigned to three FSH treatments with forty lambs in each group. 120 IU FSH was diluted by PVP with percentage 0, 15% and 30%, respectively. Meanwhile, 30% PVP treatment was subgrouped based on breed, age, hormone dose and frequency of ovum pickup, superovulation and ovum pick-up were taken in each group. In addition, in vitro fertilization and embryo transplantation of lamb oocytes were conducted compared with oocytes from slaughter house. The results showed that: 1) There were no significant differences between 30% PVP group and routine method (P>0.05), whereas 15% PVP group was lowest in each index (P<0.05); 2) There were no significant differences between Poll Dorset and Suffolk lamb in number of follicles picked per lamb and available oocytes gained per lamb (P>0.05), number of oocytes picked-up from 4, 6 and 8 week-old lambs were higher than that of 12 week-old lambs (P<0.05), when 40 IU FSH was injected, number of oocytes picked-up was lowest (P<0.05), there were no significant differences between 120 and 240 IU groups (P>0.05), efficiency of superovulation decreased significantly in second time ovum pick-up (P<0.05); 3) In vitro maturation rate, cleavage rate, morula rate and blastula rate of oocytes from lambs were lower than that of oocytes from slaughter house (P<0.05), but there was no difference in fertilization rate (P>0.05); 4) 95 and 140 blastulas from lambs and slaughter house were cryopreserved, survival rate after thawed were 95.8% and 94.3%, respectively, there were no significant differences in pregnancy rate after transplantation (P>0.05), 2 and 7 lambs were born, respectively. These results indicated that routine superovulation could be substituted by Polyvinyl pyrrolidone mediated superovulation in lambs. Number of oocytes picked-up and in vitro developmental capacity were affected by age, hormone dose and frequency of ovum pick-up, max number of available oocytes were gained when 4 week-old Poll Dorset lambs were superovulated by 120 IU FSH diluted by 30%PVP, offspring could be produced by ovum pick-up from prepubertal lambs, indicating that ovum pick-up oocytes have the same developmental capacity as oocytes from slaughter house.