Takigawa M, Danno K, Furukawa F
Department of Dermatology, Hamamatsu University School of Medicine, Japan.
Arch Dermatol Res. 1987;279(6):392-7. doi: 10.1007/BF00412625.
Regulation of the distribution of concanavalin A (Con A)/receptor complexes by the cytoskeletal contracture system was studied in guinea pig epidermal cells in suspension and culture using the fluorescence double staining method. After treatment with 100 micrograms/ml of Con A at 37 degrees C for 30 min lectin/receptor complexes were endocytosed by the less-differentiated cells in suspension and by the adherent cells in 1- and 3-day cultures that represent a growing cell fraction. The same treatment resulted in diffuse surface distribution of the complexes in the well-differentiated cells in suspension. Colchicine (10(-5) and 10(-6) M) inhibited internalization of the complexes with resultant diffuse distribution in 60% of the adherent cells in culture. Cytochalasin B (5 and 10 micrograms/ml) not only inhibited endocytosis but promoted formation of surface patchy clumps of the complexes in suspended, less-differentiated cells and cultured adherent cells. The distribution profile was not influenced by these drug treatments in the well-differentiated cells. SDS polyacrylamide gel electrophoresis and autoradiography of 125I-labelled epidermal membranes revealed several Con A-reactive polypeptides common to the cells at various differentiation steps. The progressive decrease in endocytosis and mobility of Con A/receptor complexes was suggested to occur with differentiation. In the germinative cells the distribution of lectin/receptor complexes seemed to be regulated by microfilaments and microtubules.
利用荧光双重染色法,对豚鼠悬浮培养和贴壁培养的表皮细胞中,细胞骨架挛缩系统对伴刀豆球蛋白A(Con A)/受体复合物分布的调节作用进行了研究。在37℃下用100微克/毫升的Con A处理30分钟后,悬浮培养的低分化细胞以及1天和3天贴壁培养的代表生长细胞部分的贴壁细胞内吞了凝集素/受体复合物。相同处理导致悬浮培养的高分化细胞中复合物呈弥散性表面分布。秋水仙碱(10⁻⁵和10⁻⁶摩尔)抑制了复合物的内化,导致培养的60%贴壁细胞中复合物呈弥散分布。细胞松弛素B(5和10微克/毫升)不仅抑制内吞作用,还促进悬浮培养的低分化细胞和贴壁培养细胞中复合物在表面形成斑块状聚集。在高分化细胞中,这些药物处理不影响分布情况。对125I标记的表皮膜进行SDS聚丙烯酰胺凝胶电泳和放射自显影,结果显示在不同分化阶段的细胞中存在几种与Con A反应的共同多肽。研究表明,随着细胞分化,Con A/受体复合物的内吞作用和流动性逐渐降低。在生发细胞中,凝集素/受体复合物的分布似乎受微丝和微管调节。