Selective field effects on intracellular vacuoles and vesicle membranes with nanosecond electric pulses.
Electric pulses across intact vesicles and cells can lead to transient increase in permeability of their membranes. We studied the integrity of these membranes in response to external electric pulses of high amplitude and submicrosecond duration with a primary aim of achieving selective permeabilization. These effects were examined in two separate model systems comprising of 1), a mixed population of 1,2-di-oleoyl-sn-glycero-3-phosphocholine phospholipid vesicles and in 2), single COS-7 cells, in which large endosomal membrane vacuoles were induced by stimulated endocytosis. It has been shown that large and rapidly varying external electric fields, with pulses shorter than the charging time of the outer-cell membrane, could substantially increase intracellular fields to achieve selective manipulations of intracellular organelles. The underlying principle of this earlier work is further developed and applied to the systems studied here. Under appropriate conditions, we show preferential permeabilization of one vesicle population in a mixed preparation of vesicles of similar size distribution. It is further shown that large endocytosed vacuoles in COS-7 cells can be selectively permeabilized with little effect on the integrity of outer cell membrane.
Tekle
Ephrem
E
Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892-8012, USA. ephrem@helix.nih.gov
Oubrahim
Hammou
H
Dzekunov
Sergey M
SM
Kolb
Juergen F
JF
Schoenbach
Karl H
KH
Chock
P B
PB
D000818:Animals; D019556:COS Cells; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D002463:Cell Membrane Permeability; D003593:Cytoplasm / Q000378:metabolism; D004566:Electrodes; D018274:Electroporation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008561:Membrane Fusion; D008563:Membrane Lipids; D008564:Membrane Potentials; D008566:Membranes / Q000378:metabolism; D018613:Microscopy, Confocal; D009154:Mutation; D010713:Phosphatidylcholines / Q000737:chemistry; D013997:Time Factors; D014162:Transfection; D014617:Vacuoles / Q000378:metabolism*
D008563:Membrane Lipids; D010713:Phosphatidylcholines; D049452:Green Fluorescent Proteins; C017251:1,2-oleoylphosphatidylcholine; D002118:Calcium