畜牧兽医学报 ›› 2011, Vol. 42 ›› Issue (9): 1249-1255.doi:

• 遗传繁育 • 上一篇    下一篇

利用亮甲酚蓝染色法预测牛卵母细胞发育潜能的研究

乔利敏1,李向臣4,乔富强2,姚华3,张文秀4,关伟军4*,马月辉4*   

  1. 1. 北京农业职业学院畜牧兽医系,北京 102442; 2. 北京农学院科技产业集团,北京 102206;3. 北京农学院动物科学技术学院,北京 102206; 4. 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:2011-09-23 发布日期:2011-09-23
  • 通讯作者: 关伟军,马月辉

Prediction of Bovine Oocyte Developmental Capacity by Brilliant Cresyl Blue (BCB) Staining of COCs

QIAO Li-min 1, LI Xiang-chen4, QIAO Fu-qiang2, YAO Hua3, ZHANG Wen-xiu4, GUAN Wei-jun4*, MA Yue-hui4*

  

  1. 1. Department of Animal and Veterinary, Beijing Vocational College of Agriculture, Beijing 102442, China; 2. Group of Science Technology and Industry, Beijing University of Agriculture, Beijing 102206, China; 3. Animal Science and Technology College, Beijing University of Agriculture, Beijing 102206, China; 4. Institute of Animal Science, Chinese Academy of gricultural Sciences, Beijing 100193,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-09-23 Published:2011-09-23
  • Contact: GUAN Wei-jun, MA Yuehui

摘要: 利用亮甲酚蓝(Brilliant cresyl blue,BCB)对牛未成熟卵母细胞进行染色,根据染色结果的不同对牛未成熟卵母细胞进行分组培养,观察不同组间卵母细胞发育潜能与细胞凋亡的差异,探讨以BCB着色判定牛卵质量的可行性。本试验在牛卵母细胞成熟培养前用26 μmol·L-1 BCB染色90 min作为处理组(BCB+和BCB-),以未染色卵母细胞作为对照组;以卵内谷胱甘肽的浓度作为胞质成熟的判定指标,以卵母细胞体外受精后囊胚率作为卵子发育能力的判定指标,同时参照卵母细胞的凋亡检测,综合评定卵子的质量。结果表明:(1)着色组(BCB+)卵母细胞成熟率与对照组和无色组(BCB-)间差异显著(P<0.05);(2)成熟卵母细胞中谷胱甘肽的浓度较培养前差异显著(P<0.05);(3)经BCB染色,卵母细胞在成熟后,BCB+组的成熟率、凋亡率较对照组、BCB-组差异显著(P<0.05);(4)体外受精后,BCB+组的囊胚率和囊胚细胞数较对照组和BCB-组差异显著(P<0.05)。由此可见,牛卵母细胞经BCB的染色选择可以作为判断卵母细胞未来发育潜能的一个标记。

Abstract: Bovine immature oocytes were stained by brilliant cresyl blue (BCB). Oocyte growth and apotosis between different groups was observed. The objective of the study was to investigate the feasibility of BCB staining applied to estimate bovine oocyte quality. Immature bovine oocytes were stained by 26 μmol·L-1 BCB for 90 min as experimental groups(BCB+ and BCB-) and unstained as control. Concentration of GSH in oocytes was as an index to determine cytoplasmic mature, and blastula rate was as an index to estimate oocytes growing potential after in vitro fertilization. The results showed: (1) The bovine COCs BCB+ oocyte was showed a higher mature rate than that in the BCB- and the control groups (P<0.05); (2) There was a significant difference in concentration of glutathione between mature and immature oocytes(P<0.05); (3) There was significant difference in mature rate and apoptosis rate between BCB+ group and the control and BCB- groups(P<0.05); (4) The BCB+ showed a higher blastula rate and the more blastular cells than that of BCB- and the control groups after in vitro fertilization(P<0.05). The result indicate that bovine oocyte stained by BCB could be a marker to evaluate the developmental ability of bovine oocyte in the future.