Di Lazzaro C
Medicina (Firenze). 1989 Oct-Dec;9(4):413-5.
Label-fracture technique, which allows high resolution, coincident views of immunocytochemical markers on the cell surfaces and intramembrane particles (IMPs) over platinum/carbon replicas of the exoplasmic faces of the freeze-fractured plasma membranes, was used to analyze the surface distribution and dynamics of CD3, CD4, HLA class I and II lymphocyte surface antigens, all expressed on transmembrane proteins. In prefixed cells, the native distribution of the antigens was uniform over the cell surface. Coaggregation of IMPs induced by glycerol treatment at 37 degrees C before fixation revealed codistribution of the immunolabeling and the IMPs. Redistribution of the antigens, induced by treatment at 37 degrees C (for 20 min) with specific antibodies before fixation, resulted in capping of the immunolabeling in demarcated domains of the plasma membrane but failed to reveal correspondent domains of IMPs. This observation implies that migration of antigens into the patching/capping domains may be simultaneous with the exclusion of other membrane proteins from the same areas, as occurs during viral budding and receptor mediated endocytosis.
标记断裂技术能够在冷冻断裂质膜外质面的铂/碳复制品上,对细胞表面免疫细胞化学标记物和膜内颗粒(IMPs)进行高分辨率的重合观察。该技术被用于分析CD3、CD4、HLA I类和II类淋巴细胞表面抗原的表面分布及动态变化,这些抗原均表达于跨膜蛋白上。在预先固定的细胞中,抗原在细胞表面呈均匀分布。在固定前于37℃用甘油处理诱导IMPs共聚集,结果显示免疫标记与IMPs共分布。在固定前用特异性抗体于37℃处理(20分钟)诱导抗原重新分布,导致质膜划定区域内的免疫标记形成帽状,但未能揭示相应的IMPs区域。这一观察结果表明,抗原迁移至补丁/帽状区域可能与同一区域其他膜蛋白的排除同时发生,正如病毒出芽和受体介导的内吞作用过程中所发生的那样。