Characteristics of the osmotically induced membrane rupture

The phenomenon of reciprocating mechanical oscillations of electrofused erythrocytes was used to investigate the mechanical characteristics of ruptures induced in erythrocyte membranes by colloid osmotic pressure. The rupture characteristics follow an exponentially decaying time function. Time constants determined for opening times of ruptures decreased from 5.5 ms at 10 degrees C to 3.8 ms and 2.0 ms at 40 degrees C for the first and the last observable rupture, respectively. Evidence is given that the diameter of the membrane rupture exceeds the size of a haemoglobin molecule. With repetitive membrane rupturing, the ability of the membrane bilayer and associated structures to heal decreases, owing to the reduced ability to withstand pressure gradients. This change allows oscillating doublets to be classified according to one of three groups: group A showing no development in response to swell times, group B showing a continuous decrease in response to swell times, and group C showing a spontaneous decrease in response to swell times. These results suggest that oscillations cease as a result of defects of membrane healing. Calculations of respective temperature ranges are in agreement with temperature ranges for spectrin denaturation. Thus, conclusions obtained from this study suggest that the spectrin network plays a key role in membrane healing processes after mechanical membrane rupture.