Document Type
Article
Publication Date
2014
Publication Title
Bioelectrochemistry
Volume
100
Pages
88-95
DOI
10.1016/j.bioelechem.2014.01.004
Abstract
Disruption of the actin cytoskeleton structures was reported as one of the characteristic effects of nanosecond-duration pulsed electric field (nsPEF) in both mammalian and plant cells. We utilized CHO cells that expressed the monomeric fluorescent protein (mApple) tagged to actin to test if nsPEF modifies the cell actin directly or as a consequence of cell membrane permeabilization. A train of four 600-ns pulses at 19.2 kV/cm (2 Hz) caused immediate cell membrane poration manifested by YO-PRO-1 dye uptake, gradual cell rounding and swelling. Concurrently, bright actin features were replaced by dimmer and uniform fluorescence of diffuse actin. To block the nsPEF-induced swelling, the bath buffer was isoosmotically supplemented with an electropore-impermeable solute (sucrose). A similar addition of a smaller, electropore-permeable solute (adonitol) served as a control. We demonstrated that sucrose efficiently blocked disassembly of actin features by nsPEF, whereas adonitol did not. Sucrose also attenuated bleaching of mApple-tagged actin in nsPEF-treated cells (as integrated over the cell volume), although did not fully prevent it. We conclude that disintegration of the actin cytoskeleton was a result of cell swelling, which, in turn, was caused by cell permeabilization by nsPEF and transmembrane diffusion of solutes which led to the osmotic imbalance.
Original Publication Citation
Pakhomov, A. G., Xiao, S., Pakhomova, O. N., Semenov, I., Kuipers, M. A., & Ibey, B. L. (2014). Disassembly of actin structures by nanosecond pulsed electric field is a downstream effect of cell swelling. Bioelectrochemistry, 100, 88-95. doi:10.1016/j.bioelechem.2014.01.004
Repository Citation
Pakhomov, Andrei G.; Xiao, Shu; Pakhomova, Olga N.; Semenov, Iurii; Kuipers, Marjorie A.; and Ibey, Bennett L., "Disassembly of Actin Structures by Nanosecond Pulsed Electric Field is a Downstream Effect of Cell Swelling" (2014). Bioelectrics Publications. 183.
https://digitalcommons.odu.edu/bioelectrics_pubs/183
ORCID
0000-0003-4950-4130 (Pakhomova, O.), 0000-0002-0302-1355 (Semenov)
Comments
NOTE: This is the author's pre-print version of a work that was published in Bioelectrochemistry. The final version was published as:
Pakhomov, A. G., Xiao, S., Pakhomova, O. N., Semenov, I., Kuipers, M. A., & Ibey, B. L. (2014). Disassembly of actin structures by nanosecond pulsed electric field is a downstream effect of cell swelling. Bioelectrochemistry, 100, 88-95. doi:10.1016/j.bioelechem.2014.01.004
Available at: http://dx.doi.org/10.1016/j.bioelechem.2014.01.004