Radiation-induced changes of negative charge on the cell surface of primary human fibroblasts

By binding cationized ferritin (CF) to the plasma membrane of primary human fibroblasts, the amount and topology of negatively charged sites on cell surfaces were studied after X-irradiation. The CF binding was tested both on fixed and unfixed cells. Using various enzymes, the chemical nature of sites carrying the negative charges on cell surfaces was investigated. The results suggest that in unirradiated fibroblasts the CF binding occurred in a polarized manner, i.e. the particles were localized mainly on the apical surface of cells and formed clusters. The thin cytoplasmic protrusions and cell-to-cell contact sites bound CF to a greater extent than the bleb-like formations. Enzymatic digestion of surface polysaccharides showed that the main carriers of negatively charged sites are the glycosaminoglycans associated with the cell surface. The fixation of cells with glutaraldehyde did not influence the topology of CF binding either before or after the enzymatic treatment. After X-irradiation with 2.5 Gy the topology of CF binding did not change but the CF coverage of cells as well as the amount of ferritin particles per unit of surface area decreased within 10 min. The changes proved to be reversible as the values reached the pre-irradiation level by 1 h after irradiation.