The Journal of Cell Biology, Vol 102, 1358-1362, Copyright © 1986 by The Rockefeller University Press
A long-lived fusogenic state is induced in erythrocyte ghosts by electric pulses
AE Sowers
Treatment of erythrocyte ghosts in random positions in a suspension with
membrane fusion-inducing direct current electric field pulses causes the
membranes to become fusogenic. Significant fusion yields are observed if
the membranes are dielectrophoretically aligned into membrane-membrane
contact with a weak alternating electric field as much as 5 min after the
application of the pulses. This demonstrates that a long-lived membrane
structural alteration is involved in this fusion mechanism. Other
experiments indicate that the areas on the membrane which become fusogenic
after treatment with the pulses may be very highly localized. The locations
of these fusogenic areas coincide with where the trans-membrane electric
field strength was greatest during the pulse. The fusogenic membrane
alteration, or components thereof, in these areas laterally diffuses very
slowly or not at all, or, to be fusogenic, must be present at
concentrations in the membrane above a certain threshold. The loss of
soluble 0.9-3-nm-diameter fluorescent probes from resealed cytoplasmic
compartments of randomly positioned erythrocyte ghosts occurs through
electric field pulse- induced pores only during a pulse but not between
pulses or after a train of pulses if the probe diameter is 1.2 nm or
greater. For a given pulse treatment of membranes in random positions in
suspensions, an increase in ionic strength of the medium results in (a) a
decrease in loss during the pulse, (b) no difference in loss between
pulses, and (c) an increase in fusion yield when membrane-membrane contact
is established. The latter two results (b and c) are incompatible with a
fusion mechanism that proposes a simple relationship between electric
field-induced pores and fusion.
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