Blackburn C C, Swank-Hill P, Schnaar R L
J Biol Chem. 1986 Feb 25;261(6):2873-81.
Cell surface carbohydrates and complementary carbohydrate receptors may mediate cell-cell recognition during neuronal development. To model such interactions, we developed a technique to test the ability of cell surface lipids (particularly glycosphingolipids) to mediate specific cell recognition and adhesion (Blackburn, C.C., and Schnaar, R.L. (1983) J. Biol. Chem. 258, 1180-1188). When cells were incubated on plastic microwells adsorbed with various glycolipids, carbohydrate-specific cell adhesion was readily detected. We report here the use of this method to study adhesion of embryonic chick neural retina cells to purified cell surface lipids. Rapid and specific cell adhesion was observed when the neural retina cells were incubated on surfaces adsorbed with gangliosides (an important class of neuronal cell surface glycoconjugates) but not on surfaces adsorbed with various neutral glycosphingolipids, phospholipids, or sulfatide. This suggests that the observed cell adhesion was specific for the carbohydrate moiety of the adsorbed ganglioside and was not due to nonspecific ionic or hydrophobic interactions. Although the surface density of adsorbed lipid required to support cell adhesion was the same for all gangliosides examined, the extent of adhesion varied when different purified gangliosides were used. Ganglioside-specific adhesion was not dependent on the presence of calcium (at 37 degrees C) and was attenuated by pretreatment of the cells with trypsin. The extent of ganglioside-directed neural retinal cell adhesion varied with embryonic age. These results imply that gangliosides may play a role in cell-cell recognition in the developing nervous system.
细胞表面碳水化合物和互补的碳水化合物受体可能在神经元发育过程中介导细胞间识别。为了模拟这种相互作用,我们开发了一种技术来测试细胞表面脂质(特别是糖鞘脂)介导特异性细胞识别和黏附的能力(布莱克本,C.C.,和施纳尔,R.L.(1983年)《生物化学杂志》258卷,1180 - 1188页)。当细胞在吸附有各种糖脂的塑料微孔上孵育时,很容易检测到碳水化合物特异性细胞黏附。我们在此报告使用该方法研究胚胎鸡神经视网膜细胞与纯化的细胞表面脂质的黏附。当神经视网膜细胞在吸附有神经节苷脂(一类重要的神经元细胞表面糖缀合物)的表面上孵育时,观察到快速且特异性的细胞黏附,但在吸附有各种中性糖鞘脂、磷脂或硫脂的表面上未观察到。这表明观察到的细胞黏附对吸附的神经节苷脂的碳水化合物部分具有特异性,并非由于非特异性离子或疏水相互作用。尽管对于所有检测的神经节苷脂,支持细胞黏附所需的吸附脂质表面密度相同,但使用不同纯化的神经节苷脂时,黏附程度有所不同。神经节苷脂特异性黏附不依赖于钙的存在(在37℃时),并且用胰蛋白酶预处理细胞会使其减弱。神经节苷脂介导的神经视网膜细胞黏附程度随胚胎年龄而变化。这些结果意味着神经节苷脂可能在发育中的神经系统的细胞间识别中起作用。