不同浓度β-巯基乙醇对6周龄杜泊羊卵母细胞发育能力的影响
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摘要
为了探讨成熟液里不同浓度β-巯基乙醇对6周龄杜泊羊卵母细胞发育能力的影响,试验选择6只6周龄杜泊羊,采用促性腺激素诱导活体采集卵母细胞,将获得的A、B级卵丘-卵母细胞复合体(COCs)在不同浓度(0、50、70、100μmol/L)β-巯基乙醇成熟液中成熟,再经成熟卵母细胞与获能精子共同孵育、受精卵发育培养后,统计卵裂率和囊胚率。结果表明:与对照组卵裂率(59.13%)和囊胚率(12.17%)相比,试验Ⅰ、Ⅱ、Ⅲ组的卵裂率(60.87%、63.48%、65.22%)和囊胚率(14.78%、17.39%、21.74%)均较高(P>0.05),其大小顺序为试验Ⅲ组>试验Ⅱ组>试验Ⅰ组>对照组,100μmol/Lβ-巯基乙醇组的卵裂率、囊胚率分别增加6.09个百分点和9.57个百分点。说明成熟液里添加一定量β-巯基乙醇能够提高6周龄杜泊羊卵母细胞的发育能力和受精卵质量,β-巯基乙醇浓度在100μmol/L以内,卵母细胞的卵裂率和囊胚率呈上升趋势;关于成熟液里β-巯基乙醇最佳浓度,有待进一步研究。
In order to investigate the effects of different concentrations of β-mercaptoethanol on the development ability of oocyte in Dorper sheep at the age of 6 weeks. In this experiment,6 Dorper sheep of 6-week-old were chosen to induce oocyte collection in vivo by gonadotropin,and A and B grade cumulus oocyte complexes were obtained(COCs)in different concentrations of(0,50,70,100 μmol/L)of β-mercaptoethanol in mature liquid solution. The cleavage rate and blastocyst rate were counted after matured oocytes and capacitated sperm were incubated and oosperms were cultivated. The results showed that the cleavage rates were 60. 87 %,63. 48 %,65. 22 %,blastocyst rates 14. 78 %,17. 39 %,21. 74 in group Ⅰ,Ⅱ and Ⅲ,respectively,compared with 59.13% and 12.17%(P >0.05). The order ranked test group Ⅲ>test groupⅡ>test group Ⅰ>control group,the cleavage rate and blastocyst rate of 100 μmol/L β-mercaptoethanol group increased by 6.09 and 9.57 percentage points respectively,the blastocyst rate was greater than the cleavage rate,indicating that mature liquid adding a certain amount of β-mercaptoethanol could improve the 6-week-old Dorper sheep oocyte developmental competence and the quality of fertilized ovum,beta mercaptoethanol concentration within 100 mol/L,oocyte cleavage rate and blastocyst rate would rise.The optimum concentration of beta mercaptoethanol in mature liquid needs further study.
In order to investigate the effects of different concentrations of β-mercaptoethanol on the development ability of oocyte in Dorper sheep at the age of 6 weeks. In this experiment,6 Dorper sheep of 6-week-old were chosen to induce oocyte collection in vivo by gonadotropin,and A and B grade cumulus oocyte complexes were obtained(COCs)in different concentrations of(0,50,70,100 μmol/L)of β-mercaptoethanol in mature liquid solution. The cleavage rate and blastocyst rate were counted after matured oocytes and capacitated sperm were incubated and oosperms were cultivated. The results showed that the cleavage rates were 60. 87 %,63. 48 %,65. 22 %,blastocyst rates 14. 78 %,17. 39 %,21. 74 in group Ⅰ,Ⅱ and Ⅲ,respectively,compared with 59.13% and 12.17%(P >0.05). The order ranked test group Ⅲ>test groupⅡ>test group Ⅰ>control group,the cleavage rate and blastocyst rate of 100 μmol/L β-mercaptoethanol group increased by 6.09 and 9.57 percentage points respectively,the blastocyst rate was greater than the cleavage rate,indicating that mature liquid adding a certain amount of β-mercaptoethanol could improve the 6-week-old Dorper sheep oocyte developmental competence and the quality of fertilized ovum,beta mercaptoethanol concentration within 100 mol/L,oocyte cleavage rate and blastocyst rate would rise.The optimum concentration of beta mercaptoethanol in mature liquid needs further study.
引文
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[11]马世科,乔海生,马利青,等.幼畜超数排卵及体外胚胎生产技术在青海高原地区的应用研究[J].青海畜牧兽医杂志,2015,45(2):13-14.
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[13]郭洪,万鹏程,代蓉,等.羔羊卵母细胞体外胚胎生产影响因素分析[J].湖北农业科学,2013,52(19):4726-4729.
[14]李运生,曹鸿国,刘亚,等.羔羊超数排卵与胚胎体外生产技术研究进展[J].动物医学进展,2011,32(11):99-104.
[15]韩岩,尹多,张涛,等.谷胱甘肽对卵母细胞成熟及胚胎发育影响的研究进展[J].江苏农业科学,2010(3):269-270.
[16]曹忻,赵有璋,罗海玲,等.血管内皮生长因子对体外成熟绵羊卵母细胞超微结构变化的影响[J].畜牧兽医学报,2010,41(2):167-173.
[17]郭洪,万鹏程,石文艳,等.EGF和β-巯基乙醇对羔羊卵母细胞体外受精和体外发育的影响[J].安徽农业科学,2012,40(11):6537-6540.
[18]权富生,刘琴,王丽,等.β-巯基乙醇和发情羊血清对牛颗粒细胞体外培养的影响[J].西北农业学报,2011,20(12):22-25.
[19]舒建洪,张志鹏,李向臣,等.β-巯基乙醇对牛孤雌胚胎发育的影响[J].西北农林科技大学学报,2008,36(12):24-28.
[20]兰宗宝,何若钢,莫品方,等.β-巯基乙醇对猪卵母细胞体外受精及胚胎发育的影响[J].广西畜牧兽医,2008,24(4):202-203.
[2]Trounson A O,Willadsen S M,Moor R M.Reproductive function in prepubertal lambs:ovulation,embryo development and ovarian steroidogenesis[J].J Reprod Fertil,1977,49:69-75.
[3]Kelly J M,Kleemann d O,Walker S K.Enhanced efficiency in the production of offspring from 4-to 8-week-old lambs[J].Theriogenology,2005,63(7):1876-1890.
[4]Kelly J M,Kleemann D O,Walker S K.The effect of nutrition during pregnancy on the in vitro production of embryos from resulting lambs[J].Theriogenology,2005,63(7):2020-2031.
[5]Gou K M,Guan H,Bai J H,et al.Field evaluation of juvenile in vitro embryo transfer(JIVET)in sheep[J].Animal Reproduction Science,2009,112(3/4):316-324.
[6]Bai J H,Hou J,Guan H,et al.Effect of 2-mercaptoethanol and cysteine supplementation during in vitro maturation on the developmental competence of oocytes from hormone-stimulated lambs[J].Theriogenology,2008,70(5):758-764.
[7]白佳桦.性成熟前绵羊、牛JIVET技术研究[D].北京:中国农业大学,2008.
[8]郭洪,万鹏程,石文艳,等.不同品种绵羊超数排卵及胚胎体外生产技术研究[J].畜牧与兽医,2012,44(10):20-24.
[9]汪立芹,何宗霖,林嘉鹏,等.萨福克羔羊卵泡诱导发育效果的研究[J].中国农业大学学报,2015,20(4):141-146.
[10]刘宗正,李和刚,程明,等.利用JIVET技术快速繁育萨福克羊的研究[J].畜牧与饲料科学,2016,37(3):1-3.
[11]马世科,乔海生,马利青,等.幼畜超数排卵及体外胚胎生产技术在青海高原地区的应用研究[J].青海畜牧兽医杂志,2015,45(2):13-14.
[12]陈晓勇,田树军,桑润滋,等.诱导幼羔卵泡发育及体外胚胎生产[J].农业生物技术学报,2008,16(3):456-460.
[13]郭洪,万鹏程,代蓉,等.羔羊卵母细胞体外胚胎生产影响因素分析[J].湖北农业科学,2013,52(19):4726-4729.
[14]李运生,曹鸿国,刘亚,等.羔羊超数排卵与胚胎体外生产技术研究进展[J].动物医学进展,2011,32(11):99-104.
[15]韩岩,尹多,张涛,等.谷胱甘肽对卵母细胞成熟及胚胎发育影响的研究进展[J].江苏农业科学,2010(3):269-270.
[16]曹忻,赵有璋,罗海玲,等.血管内皮生长因子对体外成熟绵羊卵母细胞超微结构变化的影响[J].畜牧兽医学报,2010,41(2):167-173.
[17]郭洪,万鹏程,石文艳,等.EGF和β-巯基乙醇对羔羊卵母细胞体外受精和体外发育的影响[J].安徽农业科学,2012,40(11):6537-6540.
[18]权富生,刘琴,王丽,等.β-巯基乙醇和发情羊血清对牛颗粒细胞体外培养的影响[J].西北农业学报,2011,20(12):22-25.
[19]舒建洪,张志鹏,李向臣,等.β-巯基乙醇对牛孤雌胚胎发育的影响[J].西北农林科技大学学报,2008,36(12):24-28.
[20]兰宗宝,何若钢,莫品方,等.β-巯基乙醇对猪卵母细胞体外受精及胚胎发育的影响[J].广西畜牧兽医,2008,24(4):202-203.