Evidence that the spectrin network and a nonosmotic force control the fusion product morphology in electrofused erythrocyte ghosts

Abstract

The conversion of the membrane area in the "contact zones" shared by erythrocyte ghosts held in contact by dielectrophoresis into a fusion product by electrofusion was studied by both light and electron microscopy. Fusion products fell into two categories: (a) those with a freely expanding open lumen which ended in the "giant cell morphology" and with considerable internal vesicle membrane fragments, and (b) linear chains of polyghosts with long term stability but having planar diaphragms at the ghost-ghost junctions. Thin section electron microscopy showed each of these planar diaphragms to be a double membrane septum multiply-perforated with fusion pores. Heat and low ionic strength treatments known to denature or detach spectrin caused the stable planar diaphragms to dissolve, thereby quickly converting the polyghost chains to the giant cell morphology, thereby suggesting that spectrin restricts fusion zone diameter expansion if it is intact. Other indications suggest that the expansion of the open lumens appears to take place as a result of one or more membrane-specific forces with a nonosmotic origin but this tendency to expansion can be overcome if the spectrin network on only one side of a contact zone is intact.