人尿促性腺激素、雌二醇及表皮生长因子对牛卵母细胞体外成熟质量的影响

doi:10.16431/j.cnki.1671-7236.2016.06.028

›› 2016, Vol. 43 ›› Issue (6): 1585-1590.doi: 10.16431/j.cnki.1671-7236.2016.06.028

人尿促性腺激素、雌二醇及表皮生长因子对牛卵母细胞体外成熟质量的影响

唐爽¹, 安文仲¹, 姜鑫龙¹, 李向臣²

1. 沈阳农业大学生物科学技术学院, 辽宁省农业生物技术重点实验室, 沈阳 110866;
2. 中国农业科学院北京畜牧兽医研究所, 国际畜禽种质资源联合实验室, 北京 100193

收稿日期:2015-10-26 出版日期:2016-06-20 发布日期:2016-07-11
通讯作者: 唐爽, 李向臣 E-mail:stfoxst@syau.edu.cn;xcli863@126.com
作者简介:唐爽(1978-),男,辽宁沈阳人,博士,讲师,研究方向:早期胚胎发育
基金资助:
辽宁省自然科学基金博士启动基金(20131104)

Effect of Human Menopausal Gonadotropin,17β-estradiol and Epidermal Growth Factor on the Quality of in vitro Maturation Bovine Oocytes

TANG Shuang¹, AN Wen-zhong¹, JIANG Xin-long¹, LI Xiang-chen²

1. Liaoning Provincial Key Laboratory for Agricultural Biotechnology, College of Biological Science and Technology, Shenyang Agricultural University, Shenyang 110866, China;
2. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2015-10-26 Online:2016-06-20 Published:2016-07-11

摘要/Abstract

摘要： 试验旨在研究不同激素配比及表皮生长因子(EGF)浓度对牛卵母细胞体外成熟及卵母细胞质量的影响。将随机分组的卵丘-卵母细胞复合体于添加FSH+LH、HMG、FSH+LH+E₂、HMG+E₂ 4种不同激素组合配比的成熟基础液中培养,对比其体外成熟率,比较了EGF对牛卵母细胞体外成熟率和孤雌胚胎体外发育的影响,并采用TUNEL法检测添加不同浓度EGF的牛孤雌激活囊胚细胞凋亡情况。结果表明,添加HMG的成熟试验结果稳定,E₂对牛卵母细胞成熟有一定的促进作用,HMG+E₂联合使用可以得到高效稳定的成熟结果;在此基础上,在成熟液中添加30 ng/mL EGF对牛卵母细胞的成熟质量、胚胎发育及降低胚胎细胞凋亡都有明显的促进作用。因此,在体外成熟培养液中添加0.075 IU/mL HMG、1 μg/mL E₂和30 ng/mL EGF对牛卵母细胞的成熟和质量较为有益。

关键词: 牛; 卵母细胞; 表皮生长因子; 人尿促性腺激素; 体外成熟

Abstract: For optimizing in vitro maturation system of bovine oocytes,we firstly examined the influence of four different hormonal regimes(FSH+LH,HMG,FSH+LH+E₂ and HMG+E₂) on oocyte maturation rates.Then we studied the effects of epidermal growth factor (EGF) in the above defined medium on bovine oocyte maturation,in vitro development and quality of parthenogenetic embryos.The cell apoptotic index of parthenogenetic blastocysts was detected by TUNEL.No significant difference was observed in maturation rates in four groups supplemented with different hormones.However,human menopausal gonadotropin (HMG) provided steady maturation results in replicates.Maturation of oocytes was promoted by supplementation with 17β-estradiol (E₂).Combination of HMG and E₂ gave rise to steady and efficient mature results.The presence of EGF at 30 ng/mL concentration significantly increased maturation rate and blastocyst rate and reduced apoptotic cells in parthenogenetic blastocysts.Therefore,the optimal oocyte maturation solution could be supplemented with 0.075 IU/mL HMG,1 μg/mL E₂ and 30 ng/mL EGF.

Key words: bovine; oocytes; epidermal growth factor; human menopausal gonadotropin; in vitro maturation

中图分类号:

Q813.7

唐爽, 安文仲, 姜鑫龙, 李向臣. 人尿促性腺激素、雌二醇及表皮生长因子对牛卵母细胞体外成熟质量的影响[J]. , 2016, 43(6): 1585-1590.

TANG Shuang, AN Wen-zhong, JIANG Xin-long, LI Xiang-chen. Effect of Human Menopausal Gonadotropin,17β-estradiol and Epidermal Growth Factor on the Quality of in vitro Maturation Bovine Oocytes[J]. , 2016, 43(6): 1585-1590.

参考文献

[1] Li R,Albertini D F.The road to maturation:somatic cell interaction and self-organization of the mammalian oocyte[J].Nat Rev Mol Cell Biol,2013,14(3):141-152.
[2] 焦明霞,刘仲凤,王峰,等.猪不同直径卵泡内卵母细胞胞质成熟及体外发育潜力差异原因的初探[J].中国畜牧兽医,2014,41(10):165-171.
[3] Gliedt D W,Rosenkrans C F,Jr Rorie R W,et al.Effects of media,serum,oviductal cells,and hormones during maturation on bovine embryo development in vitro[J].J Dairy Sci,1996,79(4):536-542.
[4] Fukui Y,Fukushima M,Terawaki Y,et al.Effect of gonadotropins,steroids and culture media on bovine oocyte maturation in vitro[J].Theriogenology,1982,18(2):161-175.
[5] Park K W,Iga K,Niwa K.Exposure of bovine oocytes to EGF during maturation allows them to develop to blastocysts in a chemically-defined medium[J].Theriogenology,1997,48(7):1127-1135.
[6] Rios G L,Buschiazzo J,Mucci N C,et al.Combined epidermal growth factor and hyaluronic acid supplementation of in vitro maturation medium and its impact on bovine oocyte proteome and competence[J].Theriogenology,2015,83(5):874-880.
[7] Beker A R,Colenbrander B,Bevers M M.Effect of 17beta-estradiol on the in vitro maturation of bovine oocytes[J].Theriogenology,2002,58(9):1663-1673.
[8] 马红,郭镇华,吴赛辉,等.不同激素对牛卵母细胞体外成熟的影响[J].中国草食动物,2010,30(2):31-33.
[9] 刘永杰,夏怡,金婧楠,等.促黄体素对牛卵母细胞体外核成熟的影响[J].中国畜牧兽医,2010,37(5):123-125.
[10] 韦精卫,韦英明,孟凡丽,等.牛卵母细胞的体外成熟[J].中国兽医学报,2008,28(2):216-219.
[11] Purohit G N,Brady M S,Sharma S S.Influence of epidermal growth factor and insulin-like growth factor 1 on nuclear maturation and fertilization of buffalo cumulus oocyte complexes in serum free media and their subsequent development in vitro[J].Anim Reprod Sci,2005,87(3-4):229-239.
[12] Lorenzo P L,Illera M J,Illera J C,et al.Enhancement of cumulus expansion and nuclear maturation during bovine oocyte maturation in vitro by the addition of epidermal growth factor and insulin-like growth factor Ⅰ[J].J Reprod Fertil,1994,101(3):697-701.
[13] Legge M.Oocyte and zygote zona pellucida permeability to macromolecules[J].J Exp Zool,1995,271(2):145-150.
[14] 李向臣,丛日华,安志兴,等.EGF和FBS对牛孤雌胚胎发育的影响[J].西北农林科技大学学报(自然科学版),2004,32(4):51-54.
[15] Reizel Y,Elbaz J,Dekel N.Sustained activity of the EGF receptor is an absolute requisite for LH-induced oocyte maturation and cumulus expansion[J].Mol Endocrinol,2010,24(2):402-411.

人尿促性腺激素、雌二醇及表皮生长因子对牛卵母细胞体外成熟质量的影响

Effect of Human Menopausal Gonadotropin,17β-estradiol and Epidermal Growth Factor on the Quality of in vitro Maturation Bovine Oocytes

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	李睿, 张伊阳, 罗雪慧, 周熳琳, 吴绍米, 杨琨, 乔自林. 低氧对牦牛肺动脉平滑肌细胞增殖及氧化应激损伤作用的影响[J]. 中国畜牧兽医, 2023, 50(6): 2245-2254.
[2]	徐媛媛, 郑海英, 杨春艳, 邓廷贤, 黄晨茜, 冯超, 陆杏蓉, 段安琴, 莫霞, 马小娅, 尚江华. 抑制BMP/Smad信号通路对水牛卵巢颗粒细胞生长和类固醇激素合成的影响[J]. 中国畜牧兽医, 2023, 50(6): 2354-2362.
[3]	肖鹏, 尚江华, 杨春艳, 李孟琪, 段安琴, 马小娅, 冯超, 黄晨茜, 张博, 周金陈, 韦科龙, 郑威, 郑海英. α-亚麻酸对水牛卵巢颗粒细胞体外培养的影响[J]. 中国畜牧兽医, 2023, 50(6): 2380-2387.
[4]	樊艺萌, 魏媛媛, 王惠茹, 尕玉, 张艳楠, 赵卿宇, 郝智慧. 基于网络药理学与分子对接探究中兽药复方乳康颗粒治疗奶牛乳房炎的作用机制[J]. 中国畜牧兽医, 2023, 50(6): 2507-2517.
[5]	张殿琦, 马鑫浩, 杨志梅, 梅楚刚, 昝林森. 秦川肉牛PAI1基因生物信息学分析及其对肌内脂肪细胞增殖和分化的影响[J]. 中国畜牧兽医, 2023, 50(5): 1729-1741.
[6]	武志娟, 柴志欣, 王吉坤, 王嘉博, 钟金城, 信金伟. 牦牛骨骼肌中特异基质金属蛋白酶及其功能相关基因的表达分析[J]. 中国畜牧兽医, 2023, 50(5): 1774-1784.
[7]	张笑梦, 焦安惠, 王玉琪, 方南洙, 金庆国. 添加罗格列酮对小鼠卵母细胞氧化还原稳态的影响[J]. 中国畜牧兽医, 2023, 50(5): 1818-1827.
[8]	彭阳翔, 杨振标, 闫奎友, 王瀚, 呼德, 王尊, 刘宁, 赵连生. 从人工到智能:牛个体识别技术研究进展[J]. 中国畜牧兽医, 2023, 50(5): 1855-1866.
[9]	黎明国, 李清春, 陈鑫, 卢世豪, 何凡, 祁梦凡, 张化鹏, 任玉军, 张庆泽, 符彬彬, 徐梦思, 艾子凯, 闫坤, 冯赟, 华再东, 黄涛, 毕延震. TET去甲基化酶对猪卵母细胞发育的影响[J]. 中国畜牧兽医, 2023, 50(5): 1907-1917.
[10]	胡湘云, 易显凤, 黄志朝, 吴思谦, 陈政谕, 吕玲燕, 黄超, 粟永春. 基于网络药理学揭示复合植物精油药理活性及作用机制[J]. 中国畜牧兽医, 2023, 50(5): 2139-2148.
[11]	胡红莲, 李超, 陈利青, 宋利文, 赵濛, 张剑霞, 王典, 高民. β-胡萝卜素对奶牛泌乳高峰期产乳性能及血常规指标、血清生化和免疫指标的影响[J]. 中国畜牧兽医, 2023, 50(4): 1375-1384.
[12]	唐毓, 杨镒峰, 张颖, 薛海龙, 冯怀亮, 许保增. 貉卵母细胞成熟发育的细胞学研究[J]. 中国畜牧兽医, 2023, 50(4): 1434-1443.
[13]	田常青, 张坤中, 齐玉梅, 史文静, 董志杰, 张浩浩, 何曾文, 芝吉, 赵学慧, 崇倩, 薛慧文, 苟惠天. 1株环境源单增李斯特菌的分离鉴定及生物特性分析[J]. 中国畜牧兽医, 2023, 50(4): 1543-1555.
[14]	和琳, 韩佃刚, 李静, 杜萱, 张家翔, 信吉阁. 牛病毒性腹泻病毒检测方法研究进展[J]. 中国畜牧兽医, 2023, 50(4): 1593-1602.
[15]	刘鑫煜, 崔强, 张兰欣, 范佺荣, 李政忆, 高瑞峰. 基于网络药理学和分子对接探究诃子鞣酸防治奶牛子宫内膜炎的作用机制[J]. 中国畜牧兽医, 2023, 50(4): 1632-1641.