外源褪黑素对牛卵母细胞体外成熟的影响

doi:10.3864/j.issn.0578-1752.2011.17.016

中国农业科学 ›› 2011, Vol. 44 ›› Issue (17): 3634-3640.doi: 10.3864/j.issn.0578-1752.2011.17.016

外源褪黑素对牛卵母细胞体外成熟的影响

高超, 田秀芝, 张璐, 徐静, 汪锋, 卓志勇, 戴蕴平, 刘国世

中国农业大学动物科技学院

收稿日期:2011-05-06 修回日期:2011-06-27 出版日期:2011-09-01 发布日期:2011-07-29
通讯作者: 通信作者刘国世，Tel：010-62732735，Fax：010-62732735；E-mail：gshliu@cau.edu.cn
作者简介:高超，E-mail：gaochao1984@yahoo.cn
基金资助:
国家“863”计划项目（2008AA1011007）、农业部转基因重大专项（2009ZX0800-020B，2008ZX08008-005）、国家科技支撑计划项目（2011BAD19B02-4）

Effect of Exogenous Melatonin (MT) in Bovine Oocyte in vitro Maturation

GAO Chao, TIAN Xiu-Zhi, ZHANG Lu, XU Jing, WANG Feng, ZHUO Zhi-Yong, DAI Yun-Ping, LIU Guo-Shi

中国农业大学动物科技学院

Received:2011-05-06 Revised:2011-06-27 Online:2011-09-01 Published:2011-07-29

摘要/Abstract

摘要： 【目的】探索外源褪黑素（melatonin，MT）处理对牛卵母细胞体外成熟的影响从而优化牛体外胚胎生产体系。【方法】在牛卵母细胞成熟过程中添加不同浓度的MT，观察并统计其体外受精后胚胎的卵裂率、囊胚率和囊胚细胞数。应用放射免疫分析法检测血清、卵泡液和成熟培养液内MT与孕酮（P）、雌激素（E2)、促卵泡素（FSH）、促黄体生成素（LH）水平的变化并检测卵母细胞的活力和活性氧（ROS）的浓度。【结果】存在于牛卵泡液中的MT浓度与血清中的相近，添加10-11 mol•L-1 MT组的P浓度（2.01±0.33）ng•mL-1显著高于对照组（（0.87±0.10）ng•mL-1，P＜0.05）。与对照组（（67.32±7.30）%）相比，10-9 mol•L-1 MT组卵裂率显著提高（（77.81±7.06）%，P＜0.05），并能提高囊胚率且囊胚细胞数（（29.61±9.55）%，（90.30±28.74）个），显著高于对照组（（19.64±9.22）%，（75.07±25.98）个，P＜0.05）；同时，10-9 mol•L-1 MT组卵母细胞的活力高于对照组，并能降低卵母细胞内的ROS含量但没有显著差异（P＞0.05）。【结论】牛卵泡液中存在MT，适当浓度的外源MT能促进牛卵母细胞体外成熟。

关键词:

褪黑素, 卵母细胞, 体外成熟, 活性氧, 牛

Abstract: 【Objective】 To optimize the bovine embryo in vitro production system，the effect of exogenous melatonin (MT) on improvement of bovine oocyte in vitro maturation was studied. 【Method】 Immature oocytes were cultured in vitro inmaturation medium supplemented with different concentrations of melatonin, then IVF statistical analysis of embryo cleavage rate, blastocyst rate and blastocyst cell number was conducted. Hormone levels were detected by radioimmunoassay in follicular fluid and the maturation medium and the the oocyte viability and reactive oxygen (ROS) levels were determined.【Result】MT existed in bovine follicular fluid and similar with serum. In 10-11 mol•L-1 treatment, the progesterone (P) levels (2.01±0.33) ng•mL-1 (P＜0.05) of maturation medium was significantly higher than that of controls (0.87±0.10) ng•mL-1 (P＜0.05). Compared with the control group (67.32±7.30)%, cleavage rate in 10-9 mol•L-1MT treatment (77.81±7.06)% was significantly increased and the blastocyst rate and cell number of blastocyst (29.61±9.55)%, (90.30±28.74) were significantly higher than that of controls (19.64±9.22)%, (75.07±25.98) (P＜0.05). 10-9 mol•L-1 MT treatment improved the oocyte viability and decreased the ROS level (P＞0.05). 【Conclusion】 Melatonin exists in bovine follicular fluid and exogenous melatonin at a proper concentration can improve the in vitro maturation of bovine oocytes.

Key words:

melatonin, oocyte, in vitro maturation, ROS, bovine

高超, 田秀芝, 张璐, 徐静, 汪锋, 卓志勇, 戴蕴平, 刘国世. 外源褪黑素对牛卵母细胞体外成熟的影响[J]. 中国农业科学, 2011, 44(17): 3634-3640.

GAO Chao, TIAN Xiu-Zhi, ZHANG Lu, XU Jing, WANG Feng, ZHUO Zhi-Yong, DAI Yun-Ping, LIU Guo-Shi. Effect of Exogenous Melatonin (MT) in Bovine Oocyte in vitro Maturation[J]. Scientia Agricultura Sinica, 2011, 44(17): 3634-3640.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2011.17.016

https://www.chinaagrisci.com/CN/Y2011/V44/I17/3634

参考文献

[1]Takasaki A, Nakamura Y, Tamura H, Shimamura K, Morioka H. Melatonin as a new drug for improving oocyte quality. Reproduction Medical Biology, 2003, 2(4): 139-144.

[2]Tamura H, Taketani A, Miwa I, Taniguchi K, Maekawa R, Asada H, Taketani T, Matsuoka A, Yamagata Y, Shimamura K, Morioka H, Ishikawa H, Reiter R J, Sugino N. Oxidative stress impairs oocyte quality and melatonin protects oocytes from radical damage and improves fertilization rate. Journal of Pineal Research, 2008, 44(3): 280-287.

[3]Voznesenskaya T, Makogon N, Bryzgina T, Sukhina V, Grushka N, Alexeyeva I. Melatonin protects against experimental immune ovarian failure in mice. Reproductive Biology, 2007, 7(3): 207-220.

[4]Kang J T, Koo O J, Kwon D K, Park H J, Jang G, Kang S K, Lee B C. Effects of melatonin on in vitro maturation of porcine oocyte and expression of melatonin receptor RNA in cumulus and granulose cells. Journal of Pineal Research, 2009, 46: 22-28.

[5]Shi J M, Tian X Z, Zhou G B, Wang L, Gao C, Zhu S E, Zeng S M, Tian J H, Liu G S. Melatonin exists in porcine follicular fluid and improves in vitro maturation and parthenogenetic development of porcine oocytes. Journal of Pineal Research, 2009, 47(4): 318-323.

[6]El-Raey M, Geshi M, Somfai T, Kaneda M, Hirako M, Abdel-Ghaffar A E, Sosa G A, Abou El-Roos M E A, Nagai T. Evidence of melatonin synthesis in the cumulus oocyte complexes and its role in enhancing oocyte maturation in vitro in cattle. Molecular Reproduction and Development, 2011, 78(4): 250-262.

[7]Manjunatha B M, Devaraj M, Gupta P S P, Ravindra J P, Nandi S. Effect of taurine and melatonin in the culture medium on buffalo in vitro embryo development. Reproduction in Domestic Animals, 2009, 44: 12-16.

[8]Brzezinski A, Seibel M M, Lynch H J, Deng M H, Wurtman R J. Melatonin in human preovulatory follicular fluid. Journal of Clinical Endocrinology and Metabolism, 1987, 64(4): 865-867.

[9]Itoh M T, Ishizuka B, Kuribayashi Y, Amemiya A, Sumi1 Y. Melatonin, its precursors and synthesizing enzyme activities in the human ovary. Molecular Human Reproduction, 1999, 5(5): 402-408.

[10]Oblap R, Olszanska B. Expression of melatonin receptor transcripts (mel-1a, mel-1b and mel-1c) in Japanese quail oocytes and eggs. Zygote, 2001, 9(3): 237-244.

[11]Papis K, Poleszczuk O, Wenta-Muchalska E, Modlinski J A. Melatonin effect on bovine embryo development in vitro in relation to oxygen concentration. Journal of Pineal Research, 2007, 43(4): 321-326.

[12]Fiske V M, Parker K L, Ulmer R A, Ow C H, Aziz N. Effect of melatonin alone or in combination with human chorionic gonadotropin or ovine luteinizing hormone on the in vitro secretion of estrogens or progesterone by granulosa cells of rats. Endocrinology, 1984, 114(2): 407-410.

[13]Webley G E, Luck M R. Melatonin directly stimulates thesecretion of progesterone by human and bovine granulosa cells luteinized in vitro. Journal of Reproduction and Fertility, 1986, 78: 711-717.

[14]Baratta M, Tamanini C. Effect of melatonin on the in vitro secretion of progesterone and estradiol 17b by ovine granulose cells. Acta Endocrinology, 1992, 127(4): 366-370.

[15]Battista P J, Condon W A. Serotonin-induced stimulation of progesterone production by cow luteal cells in vitro. Journal of Reproduction and Fertility, 1986, 76: 231-238.

[16]Brzezinski A, Fibich T, Cohen M, Shenker J G, Laufer N. Effect of melatonin on progesterone production by humangranulosa lutein cells in culture. Fertility and Sterility, 1992, 58(3): 526-529.

[17]Taniguchi K, Taketani T, Lee L M, Kizuka F, Tamura I, Sugino N. Melatonin protects granulosa cells for progesterone production as an antioxidant in human ovarian follicles. Biology of Reproduction, 2009, 81: 378.

[18]Takahashi T, Takahashi E, Igarashi H, Tezuka N, Kurachi H. Impact of oxidative stress in aged mouse oocytes on calcium oscillations at fertilization. Molecular Reproduction and Development, 2003, 66(2): 143-152.

[19]Liu L, Keefe D L. Cytoplasm mediates both development and oxidation induced apoptotic cell death in mouse zygotes. Biology of Reproduction, 2000, 62(6): 1828-1834.

[20]Tatemoto H, Sakurai N, Muto N. Protection of porcine oocytes against apoptotic cell death caused by oxidative stress during in vitro maturation: role of cumulus cells. Biology of Reproduction, 2000, 63(3): 805-810.

[21]Cetica P D, Pintos L N, Dalvit G C, Beconi M T. Antioxidant enzyme activity and oxidative stress in bovine oocyte in vitro maturation. Biochemistry and Molecular Biology International, 2001, 51(1): 57-64.

[22]Agarwal A, Saleh R A, Bedaiwy M A. Role of reactive oxygen species in the pathophysiology of human reproduction. Fertility and Sterility, 2003, 79(4): 829-843.

[23]Mouatassim S E, Guérin P, Ménézo Y. Expression of genes encoding antioxidant enzymes in human and mouse oocytes during the final stages of maturation. Molecular Human Reproduction, 1999, 5(8): 720-725.

外源褪黑素对牛卵母细胞体外成熟的影响

Effect of Exogenous Melatonin (MT) in Bovine Oocyte in vitro Maturation

PDF

赞

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0

[1]	李文慧,贺依静,姜瑶,赵红宇,彭磊,李佳,芮荣,剧世强. 伏马毒素B₁对猪体外成熟卵母细胞凋亡与自噬的影响[J]. 中国农业科学, 2022, 55(6): 1241-1252.
[2]	赫磊,路凯,赵春芳,姚姝,周丽慧,赵凌,陈涛,朱镇,赵庆勇,梁文化,王才林,朱丽,张亚东. 水稻穗顶端退化突变体paa21的表型分析及基因克隆[J]. 中国农业科学, 2022, 55(24): 4781-4792.
[3]	沙仁和,兰黎明,王三红,罗昌国. 苹果转录因子MdWRKY40b抗白粉病的机理[J]. 中国农业科学, 2021, 54(24): 5220-5229.
[4]	胡荣蓉,丁世杰,郭赟,朱浩哲,陈益春,刘政,丁希,唐长波,周光宏. Trolox对猪肌肉干细胞增殖及分化的影响[J]. 中国农业科学, 2021, 54(24): 5290-5301.
[5]	齐悦,吕峻元,张悦,韦杰,张娜,杨文香,刘大群. 小麦叶锈菌效应蛋白Pt18906激发TcLr27+31的双层防御反应[J]. 中国农业科学, 2020, 53(12): 2371-2384.
[6]	顾超珩,闫燕燕,魏夕雅,史庆华,巩彪. 硫代腺苷甲硫氨酸促进番茄百菌清降解的生理机制[J]. 中国农业科学, 2019, 52(6): 1058-1065.
[7]	张通,栗瑞兰,范晓梅,刘春洁,海日汗,霍敏,张家新. p66Shc在绵羊卵母细胞中的表达及其与胞质氧化还原稳态的关系[J]. 中国农业科学, 2019, 52(12): 2183-2192.
[8]	李海军，于泊洋，段云娇，刘星宇，翁娅政，杜晨光，王秀梅. 绵羊卵丘-卵母细胞复合体石蜡切片技术的改良[J]. 中国农业科学, 2017, 50(8): 1543-1550.
[9]	刘丰娇，蔡冰冰，孙胜楠，毕焕改，艾希珍. 富氢水浸种增强黄瓜幼苗耐冷性的作用及其生理机制[J]. 中国农业科学, 2017, 50(5): 881-889.
[10]	刘子涵，石晓艺，闫鹏娇，段阳，耿兴侠，叶佳丽，李莎，杨雪桐，张高明，贾雨林，张玲丽，宋喜悦. D²型细胞质雄性不育小麦绒毡层细胞程序化死亡与活性氧代谢[J]. 中国农业科学, 2017, 50(21): 4071-4086.
[11]	杨建军，张国斌，郁继华，胡琳莉，罗石磊，牛童，张婧. 盐胁迫下内源NO对黄瓜幼苗活性氧代谢和光合特性的影响[J]. 中国农业科学, 2017, 50(19): 3778-3788.
[12]	庞云渭，孙业清，杜卫华，郝海生，赵学明，王栋，朱化彬. 硫氧还蛋白相互作用蛋白与卵母细胞葡萄糖代谢关系的研究进展[J]. 中国农业科学, 2015, 48(S): 75-85.
[13]	乔永旭，张永平，高丽红. 根系边缘细胞对肉桂酸胁迫下黄瓜和黑籽南瓜活性氧代谢与根系活力的影响[J]. 中国农业科学, 2015, 48(8): 1579-1587.
[14]	王国骄，王嘉宇，马殿荣，苗微，赵明辉，陈温福. 不同耐冷性杂草稻和栽培稻抗氧化系统对冷水胁迫的响应[J]. 中国农业科学, 2015, 48(8): 1660-1668.
[15]	李振华，王建华. 种子活力与萌发的生理与分子机制研究进展[J]. 中国农业科学, 2015, 48(4): 646-660.