Galectins as tools for glycan mapping in histology: comparison of their binding profiles to the bovine zona pellucida by confocal laser scanning microscopy
Galectins as tools for glycan mapping in histology: comparison of their binding profiles to the bovine zona pellucida by confocal laser scanning microscopy
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DOI：
10.1007/s00418-011-0814-2
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发表时间：
2011-06-01
影响因子：
2.3
通讯作者：
Gabius, Hans-Joachim
中科院分区：
文献类型：
--
作者：
Habermann, Felix A.;Andre, Sabine;Gabius, Hans-Joachim
基因分化产生了哺乳动物凝集素的半乳糖凝集素家族。由于选择性结合不同的β-半乳糖苷是半乳糖凝集素的已知生物活性的基础，它们可以在细胞化学和组织化学中找到应用。因此，需要回答关于其各自反应性特征的相关问题。为此，需要在定义的系统中对一组半乳糖凝集素进行比较研究。在这里，我们研究了七种人凝集素以及五种源自蛋白水解截短和丝氨酸磷酸化的天然衍生物和一种工程变体的染色谱。作为试验系统，处理了具有糖蛋白包膜（透明质酸）的牛大泡囊泡卵母细胞，其呈现具有N-乙酰乳糖胺重复序列和核心岩藻糖基化的双触角至四触角复合型N-聚糖。从技术上讲，共聚焦激光扫描显微镜，首先与植物凝集素映射唾液酸化状态。因此，α 2，3/6-唾液酸化被检测到的表面丝状网状的透明，而唾液酸的聚糖链被发现覆盖的主要内部的致密层的透明。半乳糖凝集素染色是特异性的和不均匀的。在半乳糖凝集素-1至-4与半乳糖凝集素-8和-9之间，检测到表面丝状网和致密层的透明质酸的反应性的显著差异。典型的染色配置文件亲密的空间组织显示N-聚糖在不同层的透明质酸，强调潜在的半乳糖凝集素作为细胞和组织化学工具。我们的研究结果鼓励进一步的比较分析和研究，以跟踪潜在的结构和/或拓扑性质。
Gene divergence has given rise to the galectin family of mammalian lectins. Since selective binding to distinct beta-galactosides underlies the known bioactivities of galectins, they could find application in cyto- and histochemistry. The pertinent question on the characteristics of their individual reactivity profiles therefore needs to be answered. Toward this end, comparative studies of a panel of galectins in defined systems are required. We here characterise the staining profiles of seven human lectins as well as five natural derivatives originating from proteolytic truncation and serine phosphorylation and one engineered variant. As test system, bovine germinal vesicle oocytes with their glycoprotein envelope (zona pellucida), which presents bi- to tetraantennary complex-type N-glycans with N-acetyllactosamine repeats and core fucosylation, were processed. Technically, confocal laser scanning microscopy was used, first with plant lectins to map the sialylation status. Hereby, alpha 2,3/6-sialylation was detected in the superficial filamentous meshwork of the zona pellucida, while sialic acid-free glycan chains were found to characterise the main inner part of the compact layer of the zona pellucida. Galectin staining was specific and non-uniform. Significant differences in reactivity were detected for the superficial filamentous meshwork and the compact layer of the zona pellucida between galectins-1 to -4 versus galectins-8 and -9. The typical staining profiles intimate a spatially organised display of N-glycans in the different layers of the zona pellucida, underscoring the potential of galectins as cyto- and histochemical tools. Our results encourage further comparative analysis and research to trace the underlying structural and/or topological properties.