Skutelsky E, Bayer E A
J Cell Biol. 1983 Jan;96(1):184-90. doi: 10.1083/jcb.96.1.184.
The avidin-biotin complex was used for the selective ultrastructural labeling of terminal cell surface galactosyl residues. Rabbit bone marrow cells were treated with the enzyme galactose oxidase in the presence of biotin hydrazide. Subsequent treatment with ferritin-avidin conjugates enabled the electron microscopic visualization of terminal membrane-based galactose and/or N-acetylgalactosamine on these cells. All stages of erythroid development were characterized by high levels of exposed cell surface galactose, whereas all leukoid cells in the same preparations were virtually unlabeled by the above method. Modulations in the distribution of these surface determinants during differentiation and maturation of rabbit erythroid cells were found to concur in inverse fashion with respect to that of terminal sialic acids. Neuraminidase treatment, before the above labeling procedure, resulted in the exposure of additional galactosyl residues on the surface of all bone marrow cell types. The results indicate that a galactose-bearing glycoconjugate(s) may comprise an erythroid-specific membrane constituent of rabbit bone marrow cells. The high density of galactose on the surface of even the earliest erythroid precursors may eventually enable the identification and isolation of a stem cell, which already contains the erythroid-specific galactoconjugate(s). The results suggest that variations in the spectrum of cell surface carbohydrates may serve as recognition signals in the complex set of intercellular interactions which occur during the development and maturation of the erythrocyte. The occurrence of similar but species-specific variations in the complement of surface heterosaccharides during erythroid development of humans and other mammals supports this contention.
抗生物素蛋白-生物素复合物用于对终末细胞表面半乳糖基残基进行选择性超微结构标记。兔骨髓细胞在生物素酰肼存在的情况下用半乳糖氧化酶处理。随后用铁蛋白-抗生物素蛋白缀合物处理,使得能够在电子显微镜下观察这些细胞上基于膜的终末半乳糖和/或N-乙酰半乳糖胺。红细胞发育的所有阶段都以高水平暴露的细胞表面半乳糖为特征,而同一制剂中的所有白细胞实际上都未被上述方法标记。发现在兔红细胞分化和成熟过程中这些表面决定簇分布的调节与终末唾液酸的调节呈相反趋势。在上述标记程序之前用神经氨酸酶处理,导致所有骨髓细胞类型表面上额外的半乳糖基残基暴露。结果表明,一种含半乳糖的糖缀合物可能构成兔骨髓细胞的红细胞特异性膜成分。即使是最早的红细胞前体表面上半乳糖的高密度最终可能有助于鉴定和分离已经含有红细胞特异性糖缀合物的干细胞。结果表明,细胞表面碳水化合物谱的变化可能作为红细胞发育和成熟过程中发生的复杂细胞间相互作用中的识别信号。人类和其他哺乳动物红细胞发育过程中表面杂糖组成的类似但物种特异性变化的发生支持了这一论点。