Rabinovitch M, DeStefano M J
J Cell Biol. 1973 Oct;59(1):165-76. doi: 10.1083/jcb.59.1.165.
Adhesion of Sarcoma I cells (SaI) to untreated or to serum-treated glass was examined by layering (51)Cr-labeled cells on the substrate for 20 min at 34 degrees C and determining the glass-bound radioactivity after the monolayers were rinsed. Adhesion to untreated glass proceeded in sodium chloride-imidazole-potassium medium (SIK) without added divalent cations, whereas SaI adhered maximally to the serum-coated substrate only in the presence of 50 microM or more Mn. Divalent Mg, Ca, Co, Ni, or Zn were inactive or minimally active. Mn-stimulated adhesion was sharply temperature dependent, reversible upon removal of Mn, and inhibited by Ca as well as by cytochalasin B, vinblastine, or tetracaine. Adhesion of SaI in SIK did not ensue when cells or the coated substrate were pretreated with Mn and washed in SIK before the adhesion assays. Microscope observations showed that Mn induced the formation of cell processes, ruffles, and veils, and that SaI spread on the uncoated or serum-coated substrate when exposed to Mn. Cells withdrew veils and processes and rounded up when postincubated in Mn-free medium. Formation of cell processes and spreading was inhibited by cytochalasin B, vinblastine, or tetracaine. Manganese-induced adhesion seems to require the participation of microtubules and microfilaments and may be mediated by an effect of Mn on Ca fluxes. The results support the role of cell processes and spreading in cell-to-substrate adhesion.
通过在34摄氏度下将(51)铬标记的肉瘤I细胞(SaI)铺在底物上20分钟,并在冲洗单层细胞后测定玻璃结合的放射性,来检测SaI细胞与未处理或血清处理的玻璃的粘附情况。在不添加二价阳离子的氯化钠-咪唑-钾培养基(SIK)中,细胞可与未处理的玻璃粘附,而SaI细胞仅在存在50 microM或更多锰的情况下,才能最大程度地粘附于血清包被的底物。二价镁、钙、钴、镍或锌无活性或活性极低。锰刺激的粘附强烈依赖于温度,去除锰后可逆,并且受到钙以及细胞松弛素B、长春碱或丁卡因的抑制。在粘附试验前,若用锰预处理细胞或包被的底物并在SIK中洗涤,则SaI细胞在SIK中不会发生粘附。显微镜观察表明,锰可诱导细胞突起、褶皱和面纱的形成,并且当暴露于锰时,SaI细胞可在未包被或血清包被的底物上铺展。当在无锰培养基中孵育后,细胞会收起面纱和突起并变圆。细胞松弛素B、长春碱或丁卡因可抑制细胞突起的形成和铺展。锰诱导的粘附似乎需要微管和微丝的参与,并且可能是由锰对钙通量的影响介导的。这些结果支持了细胞突起和铺展在细胞与底物粘附中的作用。