Quantifying pulsed electric field-induced membrane nanoporation in single cells.

Autor: Moen EK; Ming Hsieh Department of Electrical Engineering - Electrophysics, University of Southern California, 920 Bloom Walk, SSC, 502 Los Angeles, CA, USA. Electronic address: moen.erick@gmail.com., Ibey BL; Bioeffects Division, 711 Human Performance Wing, Air Force Research Laboratory, 4141 Petroleum Rd., JBSA Fort Sam, Houston, TX 78234, USA., Beier HT; Bioeffects Division, 711 Human Performance Wing, Air Force Research Laboratory, 4141 Petroleum Rd., JBSA Fort Sam, Houston, TX 78234, USA., Armani AM; Ming Hsieh Department of Electrical Engineering - Electrophysics, University of Southern California, 920 Bloom Walk, SSC, 502 Los Angeles, CA, USA.
Jazyk: angličtina
Zdroj: Biochimica et biophysica acta [Biochim Biophys Acta] 2016 Nov; Vol. 1858 (11), pp. 2795-2803. Date of Electronic Publication: 2016 Aug 14.
DOI: 10.1016/j.bbamem.2016.08.007
Abstrakt: Plasma membrane disruption can trigger a host of cellular activities. One commonly observed type of disruption is pore formation. Molecular dynamic (MD) simulations of simplified lipid membrane structures predict that controllably disrupting the membrane via nano-scale poration may be possible with nanosecond pulsed electric fields (nsPEF). Until recently, researchers hoping to verify this hypothesis experimentally have been limited to measuring the relatively slow process of fluorescent markers diffusing across the membrane, which is indirect evidence of nanoporation that could be channel-mediated. Leveraging recent advances in nonlinear optical microscopy, we elucidate the role of pulse parameters in nsPEF-induced membrane permeabilization in live cells. Unlike previous techniques, it is able to directly observe loss of membrane order at the onset of the pulse. We also develop a complementary theoretical model that relates increasing membrane permeabilization to membrane pore density. Due to the significantly improved spatial and temporal resolution possible with our imaging method, we are able to directly compare our experimental and theoretical results. Their agreement provides substantial evidence that nanoporation does occur and that its development is dictated by the electric field distribution.
(Copyright © 2016 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE