Kinetics of sealing for transient electropores in isolated mammalian skeletal muscle cells.
Permeabilization of the plasma membrane by electrical forces (electroporation) can be either transient or stable. Although the exact molecular mechanics have not yet been described, electroporation is believed to initiate primarily in the lipid bilayer. To better understand the kinetics of membrane permeabilization, we sought to determine the time constants for spontaneous transient pore sealing. By using isolated rat flexor digitorum brevis skeletal muscle cells and a two-compartment diffusion model, we found that pore sealing times (tau p) after transient electroporation were approximately 9 min. tau p was not significantly dependent on the imposed transmembrane potential. We also determined the transmembrane potential (delta Vm) thresholds necessary for transient and stable electroporation in the skeletal muscle cells. delta VmS ranging between 340 mV and 480 mV caused a transient influx of magnesium, indicating the existence of spontaneously sealing pores. An imposed delta Vm of 540 mV or greater led to complete equilibration of the intracellular and extracellular magnesium concentrations. This finding suggests that stable pores are created by the larger imposed transmembrane potentials. These results may be useful for understanding nerve and skeletal muscle injury after an electrical shock and for developing optimal strategies for accomplishing transient electroporation, particularly for gene transfection and cell transformation.
Bier
M
M
Department of Organismal Biology and Anatomy, Pritzker School of Medicine, University of Chicago, Illinois, USA. mbier@surgery.bsd.uchicago.edu
Hammer
S M
SM
Canaday
D J
DJ
Lee
R C
RC
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002463:Cell Membrane Permeability; D018274:Electroporation / Q000379:methods*; D005260:Female; D005470:Fluorometry / Q000295:instrumentation; D066298:In Vitro Techniques; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D008564:Membrane Potentials; D018715:Microscopy, Video / Q000295:instrumentation; D008954:Models, Biological; D018482:Muscle, Skeletal / Q000166:cytology* / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D008274:Magnesium