Nanosecond electric pulses cause mitochondrial membrane permeabilization in Jurkat cells

Napotnik, Tina Batista; Wu, Yu‐Hsuan; Gundersen, M.A.; Miklavčič, Damijan; Vernier, P. Thomas

doi:10.1002/bem.20707

Cited by 144 publications

(71 citation statements)

References 45 publications

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“…These results were interpreted as a proof of a direct intracellular impact of nsEP, consistent with theoretical predictions [1,22,25,26]. However, later studies demonstrated permeabilization of the cell membrane exposed to nsEP by measuring its electrical conductance [10,15,27,28], the uptake of Tl + [6,17], Ca 2+ [29-31], water [4,10], YO-PRO-1 (YP) dye (which is smaller than Pr) [21,32], and bleomycin [33]. These data suggested that pores formed by nsEP are permeable to smaller solutes but not to Pr.…”

Section: Introductionsupporting

confidence: 68%

Multiple nanosecond electric pulses increase the number but not the size of long-lived nanopores in the cell membrane

Pakhomov

Gianulis

Vernier

et al. 2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Exposure to intense, nanosecond-duration electric pulses (nsEP) opens small but long-lived pores in the plasma membrane. We quantified the cell uptake of two membrane integrity marker dyes, YO-PRO-1 (YP) and propidium (Pr), in order to test whether the pore size is affected by the number of nsEP. The fluorescence of the dyes was calibrated against their concentrations by confocal imaging of stained homogenates of the cells. The calibrations revealed a two-phase dependence of Pr emission on the concentration (with a slower rise at < 4 µM), and a linear dependence for YP. CHO cells were exposed to nsEP trains (1 to 100 pulses, 60 ns, 13.2 kV/cm, 10 Hz) with Pr and YP in the medium, and the uptake of the dyes was monitored by time-lapse imaging for 3 min. Even a single nsEP triggered a modest but detectable entry of both dyes, which increased linearly when more pulses were applied. The influx of Pr per pulse was constant and independent of the pulse number. The influx of YP per pulse was highest with 1- and 2-pulse exposures, decreasing to about twice the Pr level for trains from 5 to 100 pulses. The constant YP/Pr influx ratio for trains of 5 to 100 pulses suggests that increasing the number of pulses permeabilizes cells to a greater extent by increasing the pore number and not the pore diameter.

show abstract

Section: Introductionsupporting

confidence: 68%

Multiple nanosecond electric pulses increase the number but not the size of long-lived nanopores in the cell membrane

Pakhomov

Gianulis

Vernier

et al. 2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

Self Cite

111

View full text Add to dashboard Cite

show abstract

“…These observations along with cellular modeling led to the conclusion that nsPEF may be preferentially permeabilizing intracellular membranes [3,9]. A recent publication supports these results by showing the uptake of barium ions into the mitochondria following nsPEF exposure [10]. Additional research focusing specifically on the plasma membrane (PM) showed that, like longer electric pulses, the PM was permeabilized by the nsPEF and remained so for minutes after the pulse, but unlike longer pulse exposures, small ions such as calcium and thallium were Distribution A.…”

Section: Introductionmentioning

confidence: 56%

Investigation of a direct effect of nanosecond pulse electric fields on mitochondria

et al. 2014

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The unique cellular response to nanosecond pulsed electric field (nsPEF) exposure, as compared to longer pulse exposure, has been theorized to be due to permeabilization of intracellular organelles including the mitochondria. In this investigation, we utilized a high-throughput oxygen and pH sensing system (Seahorse® XF24 extracellular flux analyzer) to assess the mitochondrial activity of Jurkat and U937 cells after nsPEF. The XF Analyzer uses a transient micro-chamber of only a few µL in specialized cell culture micro-plates to enable oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) to be monitored in real-time. We found that for nsPEF exposures of 10 pulses at 10-ns pulse width and at 50 kV/cm e-field, we were able to cause an increase in OCR in both U937 and Jurkat cells. We also found that high pulse numbers (>100) caused a significant decrease in OCR. Higher amplitude 150 kV/cm exposures had no effect on U937 cells and yet they had a deleterious effect on Jurkat cells, matching previously published 24 hour survival data. These results suggest that the exposures were modulating metabolic activity in cells possibly due to direct effects on the mitochondria themselves. To validate this hypothesis, we isolated mitochondria from U937 cells and exposed them similarly and found no significant change in metabolic activity for any pulse number. In a final experiment, we removed calcium from the buffer solution that the cells were exposed in and found that no significant enhancement in metabolic activity was observed. These results suggest that direct permeabilization of the mitochondria is unlikely a primary effect of nsPEF exposure and calcium-mediated intracellular pathway activation is likely responsible for observed pulse-induced mitochondrial effects.

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“…Cell permeability is the route by which the passage of different types of exogenous ions, molecules/macromolecules can pass through the cell membrane12345. This process is of substantial significance in biomedical engineering, drug delivery and gene therapy applications156789.…”

mentioning

confidence: 99%

“…This process is of substantial significance in biomedical engineering, drug delivery and gene therapy applications156789. One of the proposed causes of cell permeability is the formation of pores on the bacterial cell membrane, which is often called ‘membrane poration’12345. Cell membrane poration is known to be caused by the rearrangement of the molecular structure of the membrane, together with interactions taking place at the aqueous–lipid interface, which can be physically induced through the application of external shocks such as mechanical stress1011, ultrasound (sonoporation)2912, electric fields (electroporation)1356131415, and laser (photoporation or optoporation)416171819.…”

mentioning

confidence: 99%

18 GHz electromagnetic field induces permeability of Gram-positive cocci

Nguyen

Shamis

Croft

et al. 2015

Sci Rep

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The effect of electromagnetic field (EMF) exposures at the microwave (MW) frequency of 18 GHz, on four cocci, Planococcus maritimus KMM 3738, Staphylococcus aureus CIP 65.8T, S. aureus ATCC 25923 and S. epidermidis ATCC 14990T, was investigated. We demonstrate that exposing the bacteria to an EMF induced permeability in the bacterial membranes of all strains studied, as confirmed directly by transmission electron microscopy (TEM), and indirectly via the propidium iodide assay and the uptake of silica nanospheres. The cells remained permeable for at least nine minutes after EMF exposure. It was shown that all strains internalized 23.5 nm nanospheres, whereas the internalization of the 46.3 nm nanospheres differed amongst the bacterial strains (S. epidermidis ATCC 14990T~ 0%; Staphylococcus aureus CIP 65.8T S. aureus ATCC 25923, ~40%; Planococcus maritimus KMM 3738, ~80%). Cell viability experiments indicated that up to 84% of the cells exposed to the EMF remained viable. The morphology of the bacterial cells was not altered, as inferred from the scanning electron micrographs, however traces of leaked cytosolic fluids from the EMF exposed cells could be detected. EMF-induced permeabilization may represent an innovative, alternative cell permeability technique for applications in biomedical engineering, cell drug delivery and gene therapy.

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Nanosecond electric pulses cause mitochondrial membrane permeabilization in Jurkat cells

Cited by 144 publications

References 45 publications

Multiple nanosecond electric pulses increase the number but not the size of long-lived nanopores in the cell membrane

Multiple nanosecond electric pulses increase the number but not the size of long-lived nanopores in the cell membrane

Investigation of a direct effect of nanosecond pulse electric fields on mitochondria

18 GHz electromagnetic field induces permeability of Gram-positive cocci

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