Gadolinium induces domain and pore formation of human erythrocyte membrane: an atomic force microscopic study

Lanthanide cations bind to human erythrocyte membranes and enhance cell permeability. It was postulated that this effect is due to their likeness with calcium ions, which have been used to induce perforation of cells. However, the nature and mechanism of the perforation are still not clear. In the present work, the change in surface topography of erythrocyte membranes exposed to various gadolinium species was imaged with an atomic force microscope (AFM) in order to get direct evidence of perforation. The images of the whole cell and regions in nanometer scale showed that the normal surface is featured by closely packed nanometer size particles. The AFM images showed that Gd(3+) binding to erythrocytes led to domain structure at low concentration and pore formation at higher concentration. The domain structures that appeared after incubation with 1.0x10(-6)-1.0x10(-5) mol/l Gd(3+) solution for 30 min are featured by the particles aggregated to form ranges and the separations among them enlarged to gorges. With a higher concentration, 2.5x10(-5) mol/l Gd(3+), the further aggregation developed into crater-shaped 'pores'. By washing with EDTA the 'pores' can be resealed but the domain structure remained. The anionic complex of Gd(3+), [Gd(Cit)(2)](3-) of this concentration, can only induce the domain structure formation. The domain and 'pore' structures mediated by Gd(3+) concentrations might be responsible for both enhanced permeability and perforation. The mechanism of Gd-induced domain formation and perforation is discussed on the basis of aggregation of membrane proteins and the coexistence of different phases of membrane lipids resulting from Gd(3+) binding.