Electropermeabilization of endocytotic vesicles in B16 F1 mouse melanoma cells

It has been reported previously that electric pulses of sufficiently high voltage and short duration can permeabilize the membranes of various organelles inside living cells. In this article, we describe electropermeabili- zation of endocytotic vesicles in B16 F1 mouse melanoma cells. The cells were exposed to short, high-voltage electric pulses (from 1 to 20 pulses, 60 ns, 50 kV/cm, repetition frequency 1 kHz). We observed that 10 and 20 such pulses induced permeabilization of membranes of endocytotic vesicles, detected by release of lucifer yellow from the vesicles into the cytosol. Simultaneously, we detected uptake of propidium iodide through plasma membrane in the same cells. With higher number of pulses permeabilization of the membranes of endocytotic vesicles by pulses of given parameters is accompanied by permeabilization of plasma membrane. However, with lower number of pulses only permeabilization of the plasma membrane was detected.

[1]  M C Willingham,et al.  Modified LDLs are internalized by macrophages in part via macropinocytosis , 1999, The Anatomical record.

[2]  J. Teissié,et al.  Ionic modulation of electrically induced fusion of mammalian cells , 2005, The Journal of Membrane Biology.

[3]  H. McMahon,et al.  Snap-shots of clathrin-mediated endocytosis. , 2002, Trends in biochemical sciences.

[4]  K. Schoenbach,et al.  Submicrosecond intense pulsed electric field effects on intracellular free calcium: mechanisms and effects , 2004, IEEE Transactions on Plasma Science.

[5]  E. Neumann,et al.  Electroporation and Electrofusion in Cell Biology , 1989, Springer US.

[6]  J. Swanson,et al.  M-CSF-induced macropinocytosis increases solute endocytosis but not receptor-mediated endocytosis in mouse macrophages. , 1992, Journal of cell science.

[7]  Damijan Miklavcic,et al.  The effect of electroporation pulses on functioning of the heart , 2008, Medical & Biological Engineering & Computing.

[8]  Juergen F Kolb,et al.  Membrane permeabilization and cell damage by ultrashort electric field shocks. , 2007, Archives of biochemistry and biophysics.

[9]  Laura Marcu,et al.  Nanoelectropulse-induced phosphatidylserine translocation. , 2004, Biophysical journal.

[10]  Laura Marcu,et al.  Pulse generators for pulsed electric field exposure of biological cells and tissues , 2003 .

[11]  U. Pliquett,et al.  Joule heating during solid tissue electroporation , 2003, Medical and Biological Engineering and Computing.

[12]  T M Jovin,et al.  Fluorescence labeling and microscopy of DNA. , 1989, Methods in cell biology.

[13]  I. Ferguson,et al.  Major changes in chromatin condensation suggest the presence of an apoptotic pathway in plant cells. , 1998, Experimental cell research.

[14]  K. Schoenbach,et al.  Nanosecond pulsed electric fields modulate cell function through intracellular signal transduction mechanisms. , 2004, Physiological measurement.

[15]  Juergen F Kolb,et al.  Nanosecond pulsed electric field generators for the study of subcellular effects , 2006, Bioelectromagnetics.

[16]  A. Gandolfi,et al.  Study of propidium iodide binding to DNA in intact cells by flow cytometry , 2008, Cell Biophysics.

[17]  Steve W. Smye,et al.  Membrane electroporation theories: a review , 2006, Medical and Biological Engineering and Computing.

[18]  Ravindra P. Joshi,et al.  Ultrashort electrical pulses open a new gateway into biological cells , 2004, Proceedings of the IEEE.

[19]  K. Schoenbach,et al.  Simulations of nanopore formation and phosphatidylserine externalization in lipid membranes subjected to a high-intensity, ultrashort electric pulse. , 2005, Physical review. E, Statistical, nonlinear, and soft matter physics.

[20]  Robert G Parton,et al.  Clathrin-independent endocytosis: new insights into caveolae and non-caveolar lipid raft carriers. , 2005, Biochimica et biophysica acta.

[21]  M Cemazar,et al.  Electrically-assisted nucleic acids delivery to tissues in vivo: where do we stand? , 2006, Current pharmaceutical design.

[22]  K. Schoenbach,et al.  Nanosecond, high‐intensity pulsed electric fields induce apoptosis in human cells , 2003, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[23]  J. Weaver,et al.  Theory of electroporation: A review , 1996 .

[24]  Juergen F Kolb,et al.  Selective field effects on intracellular vacuoles and vesicle membranes with nanosecond electric pulses. , 2005, Biophysical journal.

[25]  A. T. Esser,et al.  Microdosimetry for conventional and supra-electroporation in cells with organelles. , 2006, Biochemical and biophysical research communications.

[26]  Ludger Johannes,et al.  Clathrin‐Dependent or Not: Is It Still the Question? , 2002, Traffic.

[27]  Robert Langer,et al.  Transdermal drug delivery , 2008, Nature Biotechnology.

[28]  Martin A Gundersen,et al.  Nanosecond electric pulse-induced calcium entry into chromaffin cells. , 2008, Bioelectrochemistry.

[29]  K. Schoenbach,et al.  Nanosecond pulsed electric field (nsPEF) effects on cells and tissues: apoptosis induction and tumor growth inhibition , 2001, PPPS-2001 Pulsed Power Plasma Science 2001. 28th IEEE International Conference on Plasma Science and 13th IEEE International Pulsed Power Conference. Digest of Papers (Cat. No.01CH37251).

[30]  Damijan Miklavcic,et al.  Optimization of bulk cell electrofusion in vitro for production of human-mouse heterohybridoma cells. , 2008, Bioelectrochemistry.

[31]  D Miklavcic,et al.  The effect of pulse repetition frequency on the uptake into electropermeabilized cells in vitro with possible applications in electrochemotherapy. , 2002, Bioelectrochemistry.

[32]  John H Ashmore,et al.  Characterization of the cytotoxic effect of high-intensity, 10-ns duration electrical pulses , 2004, IEEE Transactions on Plasma Science.

[33]  U. Pliquett,et al.  Nanosecond pulsed electric fields cause melanomas to self-destruct , 2006, Conference Record of the 2006 Twenty-Seventh International Power Modulator Symposium.

[34]  James C Weaver,et al.  Electrical behavior and pore accumulation in a multicellular model for conventional and supra-electroporation. , 2006, Biochemical and biophysical research communications.

[35]  Karl H. Schoenbach,et al.  Nanosecond pulsed electric fields induce apoptosis in p53-wildtype and p53-null HCT116 colon carcinoma cells , 2007, Apoptosis.

[36]  Damijan Miklavcic,et al.  Blumlein Configuration for High-Repetition-Rate Pulse Generation of Variable Duration and Polarity Using Synchronized Switch Control , 2009, IEEE Transactions on Biomedical Engineering.

[37]  K. Schoenbach,et al.  Intracellular effect of ultrashort electrical pulses , 2001, Bioelectromagnetics.

[38]  Juergen F Kolb,et al.  Long‐lasting plasma membrane permeabilization in mammalian cells by nanosecond pulsed electric field (nsPEF) , 2007, Bioelectromagnetics.

[39]  Juergen F Kolb,et al.  Nanosecond electric pulses penetrate the nucleus and enhance speckle formation. , 2007, Biochemical and biophysical research communications.

[40]  K. Schoenbach,et al.  Diverse effects of nanosecond pulsed electric fields on cells and tissues. , 2003, DNA and cell biology.

[41]  Shu Xiao,et al.  Nanosecond pulse electric field (nanopulse): a novel non-ligand agonist for platelet activation. , 2008, Archives of biochemistry and biophysics.

[42]  Laura Marcu,et al.  Nanosecond pulsed electric fields perturb membrane phospholipids in T lymphoblasts , 2004, FEBS letters.

[43]  Martin A Gundersen,et al.  Nanopore-facilitated, voltage-driven phosphatidylserine translocation in lipid bilayers—in cells and in silico , 2006, Physical biology.

[44]  Laura Marcu,et al.  In vitro and in vivo evaluation and a case report of intense nanosecond pulsed electric field as a local therapy for human malignancies , 2007, International journal of cancer.

[45]  Tatsuo Yamada,et al.  Amyloid-beta causes apoptosis of neuronal cells via caspase cascade, which can be prevented by amyloid-beta-derived short peptides , 2005, Experimental Neurology.

[46]  D Miklavcic,et al.  Electrochemotherapy in treatment of tumours. , 2008, European journal of surgical oncology : the journal of the European Society of Surgical Oncology and the British Association of Surgical Oncology.

[47]  Damijan Miklavčič,et al.  Cell membrane fluidity related to electroporation and resealing , 2006, European Biophysics Journal.

[48]  Laura Marcu,et al.  Calcium bursts induced by nanosecond electric pulses. , 2003, Biochemical and biophysical research communications.

[49]  Damijan Miklavcic,et al.  Theoretical evaluation of voltage inducement on internal membranes of biological cells exposed to electric fields. , 2006, Biophysical journal.

[50]  Juergen F Kolb,et al.  Leukemic cell intracellular responses to nanosecond electric fields. , 2004, Biochemical and biophysical research communications.

[51]  Michael R Murphy,et al.  Plasma membrane permeabilization by 60‐ and 600‐ns electric pulses is determined by the absorbed dose , 2009, Bioelectromagnetics.

[52]  Boris Rubinsky,et al.  Tumor Ablation with Irreversible Electroporation , 2007, PloS one.

[53]  D. Miklavčič,et al.  Electropermeabilization of dense cell suspensions , 2007, European Biophysics Journal.

[54]  Sandra L. Schmid,et al.  Regulated portals of entry into the cell , 2003, Nature.

[55]  R. O. Price,et al.  Plasma membrane voltage changes during nanosecond pulsed electric field exposure. , 2006, Biophysical journal.

[56]  Takayuki Oshima,et al.  Bacterial sterilization and intracellular protein release by a pulsed electric field. , 2004, Advances in biochemical engineering/biotechnology.

[57]  David Owen,et al.  Life of a clathrin coat: insights from clathrin and AP structures , 2006, Nature Reviews Molecular Cell Biology.

[58]  Martin A Gundersen,et al.  Nanoelectropulse-driven membrane perturbation and small molecule permeabilization , 2006, BMC Cell Biology.

[59]  K. Schoenbach,et al.  Differential effects in cells exposed to ultra-short, high intensity electric fields: cell survival, DNA damage, and cell cycle analysis. , 2003, Mutation research.

[60]  J. Hancock,et al.  Lipid rafts and membrane traffic , 2007, FEBS letters.