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Post-Fusion Membrane Reorganization.
Final rept. 28 Oct 91-31 Jan 93,
MARYLAND UNIV BALTIMORE SCHOOL OF MEDICINE
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We have previously shown by thin section electron microscopy BJ 60 1037 in a contact zone induced between erythrocyte ghosts by dielectrophoresis, the use of an electrofusion protocol converts the contact zone into a fusion zone FZ containing from 1-225 fusion pores per micrometer squared. Also, after creation, both the FZ stability, and the time-dependent diameter expansion rates of the fusion zone are strongly dependent on an intact spectrin network. In the present project, and using computer-assisted analysis on video-recorded phase optics images of fusion zone diameters, we have found that the FZ diameter vs. time kinetics has two, or three, distinct phases I-III in erythrocyte ghosts with an intact or heat-disrupted 42 oC, 20 min, respectively, spectrin network. These phases, revealed by FZ diameter vs. time measurements under some combinations of various electric pulse parameters, temperatures 4-33 deg C during measurements, heat treatment temperatures 39-50 deg C, and dielectrophoretic force E sub ac 3.25, 4.25, and 5.25 Vmm, and under certain conditions, were remarkably independent from one another, yet the durations of Phase I 1.0-1.2 sec and Phase II 4.0 sec were remarkably invariant regardless of the variable studied. This suggested the existence of a complex but dissectable interplay of biomechanical factors....Membranes, Cells, Electrofusion, Electroporation, Red cells, Erythrocytes.
APPROVED FOR PUBLIC RELEASE