中国农业科学 ›› 2007, Vol. 40 ›› Issue (8): 1780-1785 .

• 畜牧·兽医·资源昆虫 • 上一篇    下一篇

一个重要的精子受体—猪透明带糖蛋白β-D-半乳糖残基

宋学雄,曹国强,张进玉,张 圆,于晓云   

  1. 青岛农业大学动物科技学院
  • 收稿日期:2005-12-21 修回日期:1900-01-01 出版日期:2007-08-10 发布日期:2007-08-10

An important sperm receptor—beta-D-galactosyl residue of Zona pellucida glycoprotein in pig

  1. 青岛农业大学动物科技学院
  • Received:2005-12-21 Revised:1900-01-01 Online:2007-08-10 Published:2007-08-10

摘要: 【目的】证明末端半乳糖残基在猪透明带糖蛋白中的分布以及它在精子结合和侵入中的作用。【方法】体外成熟的猪卵母细胞去除周围卵丘细胞,用异硫氰酸荧光素(fluorescein isothiocyanate,FITC) 络合西非单叶豆凝集素(Bandeiraea simplicifolia lectin-I,BS-I) 或蓖麻凝集素(Ricinus communis agglutinin,RCA-I)处理30min,分别观察末端-D-半乳糖残基或-D-半乳糖残基在透明带中的定位分布。荧光显微镜观察显示,BS-I和RCA-I都标记于整个透明带上且比较集中于透明带的外层,其中BS-I显示微弱荧光反应,而RCA-I呈强荧光反应且在透明带的外层反应尤为强烈。植物凝集素处理的卵母细胞用于体外受精,分别在受精2h或12h观察精-卵结合和精子入卵情况。【结果】RCA-I处理组卵母细胞平均精子结合数显著低于对照组(18 vs 95),而精子入卵则完全被阻断;可是BS-I处理组的卵母细胞的精子结合数少量被抑制(59 vs 95),而且精子入卵率与对照组无显著差异。【结论】猪透明带末端-D-半乳糖残基作为精子受体的重要组成成分,参与精-卵结合和受精。

关键词: 精子受体, 猪透明带, 糖蛋白, β-D-半乳糖残基, 植物凝集素

Abstract: Abstract: The present study aimed to identify the distribution of D-galactosyl residue on zona pellucida (ZP) and its function(s) in sperm binding and penetration in vitro in pig. In vitro matured pig oocytes were freed from cumulus cells and treated for 30 mins with fluorescein isothiocyanate(FITC)-labeled lectin Bandeiraea simplicifolia lectin-I (BS-I) and Ricinus communis agglutinin (RCA-I) to examine alpha-D-galactosyl residue and beta-D-galactosyl residue respectively. When examined under a fluorescence microscope, both BS-I and RCA-I were bound to the ZP and most of them accumulated in the outer surface of the ZP. Among them, bound BS-I showed weaker fluorescence but bound RCA-I showed stronger fluorescence especially in the outer surface of the ZP. The treated oocytes were inseminated in vitro and after 2 or 12h insemination, the sperm-oocyte binding and penetration were examined. The results showed that after treatment of RCA-I, the sperm binding number was inhibited (18 vs 95) and sperm penetration was completely blocked; after treatment of BS-I, the sperm binding number was slightly inhibited (59 vs 95) and sperm penetration was not influenced. These results suggested that beta-D-galactosyl residue in the pig ZP glycoprotein may act as an important sperm receptor for sperm-oocyte binding and penetration.

Key words: sperm receptor, pig zona pellucida, glycoprotein, beta-D-galactosyl residues, Lectin