Spectrin, red cell shape and deformability

Summary

Using novel microscopic techniques for observing individual cells in suspension, the shape and deformability of the erythrocytes of a spectrin-deficient strain of mouse were investigated in vitro (fresh and after fixation) and in vivo (intravital microscopy of the mesenteric capillaries). The animals were identical to those studied by Greenquist et al. [14]; however, in contrast to the descriptions by these authors, in the present study, spherical cells were seen only exceptionally. Instead, sphero-echinocytes, spherules with tethered microvesicles, myelin figures and occasional stomato-spherocytes were observed. This pleomorphy, also seen in scanning electron micrographs, can be explained by the fact that in the majority of these cells the membrane assumes an extremely positive outward curvature. After osmotic dehydration, all cells responded by developing progressive membrane protrusions with a positive curvature. Osmotic inflation led to reincorporation of the membrane tethers. After hypoosmolar lysis, the ghosts of the spectrin-deficient mouse cells also showed a pronounced tendency to assume shapes characterized by a progressively positive outward curvature. In vivo observation of the mouse erythrocytes using high magnification interference contrast optics confirmed this in vitro observation. Thus, it is concluded that spectrin deficiency does not primarily lead to spherocytosis: instead, the equilibrium shapes assumed are associated with membrane evaginations which are easily torn off from the main cell body by mechanical forces.