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用凝集素检测豚鼠活体椭圆囊斑中的碳水化合物分布。

Carbohydrate distribution in the living utricular macula of the guinea pig detected by lectins.

作者信息

Suzuki H, Katori Y, Ikeda K, Takasaka T

机构信息

Department of Otolaryngology, Tohoku University School of Medicine, Sendai, Japan.

出版信息

Hear Res. 1995 Jul;87(1-2):32-40. doi: 10.1016/0378-5955(95)00075-f.

Abstract

Carbohydrate distribution in the fresh utricular macula of the guinea pig was analysed using lectins such as Concanavalin A (ConA), Dolichos biflorus agglutinin (DBA), peanut agglutinin (PNA), soybean agglutinin (SBA), Ulex europeus agglutinin (UEA-1) and wheat germ agglutinin (WGA) by means of confocal laser scanning microscopy. The ciliary bundle was strongly reactive to ConA, PNA, SBA and WGA but not to DBA and UEA-I, showing that the ciliary bundle has abundant D-galactose (GaI), N-acetyl-D-glucosamine (GlcNAc), D-mannose (Man) and sialic acid(s) (Sia) but not detectable amounts of L-fucose (Fuc) and terminal N-acetyl-D-galactosamine (GalNAc). Similar patterns of lectin bindings with moderate-to-weak intensities were observed on the non-cilial apical surface, on the surface of the otoconia and in the gelatinous layer of the otoconial membrane. On the contrary, the globular substance, a precursor of the otoconia, was scarcely reactive to any lectin examined, implying that it lacks glycoconjugates on its surface. Previous histochemical studies reported that the otoconial membrane possesses a much higher affinity for lectins that does the sensory epithelium (including the cilia) in the vestibular organ. This discrepancy suggests that factors in the preparation process may affect the otoconial membrane or the surface coat of the cilia to change their lectin affinity. Meanwhile, sialidase treatment augmented the affinity of the ciliary bundle for DBA and PNA, indicating that sialylated GalNAc and Gal are present on the vestibular ciliary bundle.

摘要

通过共聚焦激光扫描显微镜,使用诸如刀豆球蛋白A(ConA)、双花扁豆凝集素(DBA)、花生凝集素(PNA)、大豆凝集素(SBA)、荆豆凝集素(UEA-1)和小麦胚芽凝集素(WGA)等凝集素,分析豚鼠新鲜椭圆囊斑中的碳水化合物分布。纤毛束对ConA、PNA、SBA和WGA有强烈反应,但对DBA和UEA-I无反应,表明纤毛束含有丰富的D-半乳糖(Gal)、N-乙酰-D-葡萄糖胺(GlcNAc)、D-甘露糖(Man)和唾液酸(Sia),但未检测到L-岩藻糖(Fuc)和末端N-乙酰-D-半乳糖胺(GalNAc)。在非纤毛顶端表面、耳石表面和耳石膜的凝胶层上观察到类似的凝集素结合模式,强度为中度至弱度。相反,耳石的前体球状物质对所检测的任何凝集素几乎没有反应,这意味着其表面缺乏糖缀合物。先前的组织化学研究报道,耳石膜对凝集素的亲和力比前庭器官中的感觉上皮(包括纤毛)高得多。这种差异表明,制备过程中的因素可能会影响耳石膜或纤毛的表面涂层,从而改变它们对凝集素的亲和力。同时,唾液酸酶处理增强了纤毛束对DBA和PNA的亲和力,表明前庭纤毛束上存在唾液酸化的GalNAc和Gal。

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