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大鼠卵子透明带糖蛋白的受精后变化

Post-fertilization changes in the zona pellucida glycoproteins of rat eggs.

作者信息

Raz T, Skutelsky E, Shalgi R

机构信息

Department of Embryology, Sackler School of Medicine, Tel Aviv University, Ramat Aviv, Israel.

出版信息

Histochem Cell Biol. 1996 Oct;106(4):395-403. doi: 10.1007/BF02473298.

Abstract

The zona pellucida (ZP) is the extracellular coat surrounding the mammalian egg. Numerous evidence supports the role of ZP carbohydrate residues as the specific sperm receptors. In this study we used lectins to study different distribution patterns of carbohydrate residues in the rat ZP, and to follow changes at fertilization. ZP were collected from follicular, ovulated, and fertilized eggs, incubated with one of 11 different biotin-labeled lectins, followed by avidin-fluorescein isothiocyanate (FITC) complex, and visualized by epifluorescent microscopy. For electron microscope (EM) histochemistry, eggs were embedded in LR white and ultrathin sections were stained with the complex Ricinus communis lectin (RCA-1)-colloidal gold. Some lectins (RCA-I, Glycine max) bound to the entire ZP while others were restricted to the inner or outer zones [Griffonia simplicifolia, Concanovalia ensiformis, Triticum vulgaris (WGA), succinyl-WGA]. Other lectins (Lens culinaris, Ulex europhaeus) were totally excluded. The RCA-1 binding pattern changed following sperm penetration, from homogeneous in ZP of ovulated eggs (57%) to uneven in ZP of fertilized (71%) or activated (68%) eggs. Our results demonstrate an uneven distribution of different sugar residues in the rat ZP, and a post-fertilization change in the distribution of beta-galactose, which is specifically recognized by RCA-I, presumably correlated with other changes in the ZP that lead to the block to polyspermy.

摘要

透明带(ZP)是围绕哺乳动物卵子的细胞外被膜。大量证据支持ZP碳水化合物残基作为特定精子受体的作用。在本研究中,我们使用凝集素来研究大鼠ZP中碳水化合物残基的不同分布模式,并追踪受精时的变化。从卵泡期、排卵后和受精后的卵子中收集ZP,与11种不同的生物素标记凝集素之一孵育,然后与抗生物素蛋白-异硫氰酸荧光素(FITC)复合物结合,并通过落射荧光显微镜观察。对于电子显微镜(EM)组织化学,将卵子包埋在LR白色树脂中,超薄切片用蓖麻凝集素(RCA-1)-胶体金复合物染色。一些凝集素(RCA-I、大豆凝集素)与整个ZP结合,而其他凝集素则局限于内部或外部区域[简叶相思子凝集素、刀豆凝集素、小麦胚芽凝集素(WGA)、琥珀酰-WGA]。其他凝集素(小扁豆凝集素、荆豆凝集素)则完全不结合。精子穿透后,RCA-1的结合模式发生变化,从排卵后卵子ZP中的均匀分布(57%)变为受精(71%)或激活(68%)卵子ZP中的不均匀分布。我们的结果表明大鼠ZP中不同糖残基分布不均,受精后β-半乳糖分布发生变化,RCA-I能特异性识别β-半乳糖,推测这与ZP中导致多精受精阻断的其他变化相关。

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