Carbohydrate-specific cell adhesion directly to glycosphingolipids separated on thin-layer chromatography plates.
Carbohydrate-specific cell adhesion directly to glycosphingolipids separated on thin-layer chromatography plates.
复制标题
碳水化合物特异性细胞直接粘附到薄层色谱板上分离的鞘糖脂上。
DOI：
10.1016/0003-2697(87)90088-1
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发表时间：
1987
影响因子：
2.9
通讯作者：
Schnaar,RL
中科院分区：
文献类型：
--
作者：
Swank-Hill,P;Needham,LK;Schnaar,RL
细胞表面糖和互补糖受体可介导细胞-细胞识别和粘附。我们报告了一种方法，检测完整的真核细胞直接糖鞘脂分离薄层层析板上的碳水化合物特异性粘附。在高效硅胶薄层色谱板上对各种鞘糖脂进行色谱分析，并将板用聚（甲基丙烯酸异丁酯）薄膜涂覆并安装在专门设计的有机玻璃室中。将代谢性放射性标记的细胞加入到腔室中，然后密封并轻轻离心，使细胞与TLC板表面接触。孵育以使粘附发生后，将腔室倒置并离心以从板表面去除未粘附的细胞。将平板从室中取出，用戊二醛将粘附细胞固定在适当位置，干燥平板并进行放射自显影。鸡肝细胞具有N-乙酰葡糖胺的细胞表面受体，仅粘附于适当鞘糖脂（具有该末端糖）迁移的平板区域。细胞粘附被可溶性N-乙酰葡糖胺阻断（但不被其他糖阻断），并且使用各种显影溶剂很容易检测到。很容易检测到细胞粘附到低至8 pmol的适当脂质。该方法可用于测试作为多种细胞类型的细胞表面识别标记物的鞘糖脂。
Cell surface carbohydrates and complementary carbohydrate receptors may mediate cell-cell recognition and adhesion. We report a method which detects carbohydrate-specific adhesion of intact eukaryotic cells directly to glycosphingolipids separated on thin-layer chromatography plates. Various glycosphingolipids were chromatographed on high-performance silica gel thinlayer chromatography plates, and the plates were coated with a thin film of poly(isobutyl methacrylate) and mounted in a specially designed plexiglass chamber. Metabolically radiolabeled cells were added to the chamber, which was then sealed and gently centrifuged to bring the cells into contact with the surface of the TLC plate. After incubation to allow adhesion to occur, the chamber was inverted and centrifuged to remove nonadherent cells from the plate surface. The plate was removed from the chamber, the adherent cells were fixed in place with glutaraldehyde, and the plate was dried and subjected to autoradiography. Chicken hepatocytes, which have a cell surface receptor for N-acetylglucosamine, adhered only to those areas of the plate to which appropriate glycosphingolipids (having that terminal sugar) had migrated. Cell adhesion was blocked by soluble N-acetylglucosamine (but not by other sugars) and was readily detectable using a variety of developing solvents. Cell adhesion to as little as 8 pmol of the appropriate lipid was readily detected. This method can be used to test glycosphingolipids as cell surface recognition markers for a variety of cell types.