Abstract
An exposure of a cell to an external electric field results in the induced transmembrane voltage (ΔΨm) that superimposes to the resting voltage. This can have a range of effects, from modification of the activity of voltage-gated channels to membrane electroporation, and accurate knowledge of spatial distribution and time course of ΔΨm is important for the understanding of these effects. In this chapter, we present the analytical, numerical, and experimental methods of determination of ΔΨm, and combine them with the monitoring of electroporation-induced transmembrane molecular transport (TMT) in Chinese Hamster Ovary (CHO) cells. Potentiometric measurements are performed using di-8-ANEPPS, and TMT is monitored using propidium iodide. In isolated cells, we combine analytical derivation (for spherical cells) and numerical computation of ΔΨm (for irregularly shaped cells) with potentiometric measurements to show that the latter are accurate and reliable. Monitoring of TMT in these same cells shows that it is confined to the regions with the highest |ΔΨm|. We then review other parameters influencing electroporation of isolated cells, and proceed, through the intermediate case of dense suspensions, to cells in direct contact with each other. We use the scrape-loading test to show that the CHO cells in a monolayer are interconnected, and then study ΔΨm and TMT in a cluster of four such cells. With low pulse amplitudes, the cluster behaves as one big cell, with ΔΨm continuous along its outer boundary, reflecting the interconnections. With interconnections inhibited, the cells start to behave as individual entities, with ΔΨm continuous along the plasma membrane of each cell. With the cluster exposed to porating (higher amplitude) pulses, TMT occurs in the membrane regions for which computations predict the highest |ΔΨm| if the cells are modeled as insulated, suggesting that the interconnections are blocked by supraphysiological ΔΨm, either directly by voltage gating or indirectly through changes in ionic concentrations caused by electroporation.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bedlack RS, Wei M, Fox SH, et al. Distinct electric potentials in soma and neurite membranes. Neuron. 1994;13:1187–93.
Cheng DKL, Tung L, Sobie EA. Nonuniform responses of transmembrane potential during electric field stimulation of single cardiac cells. Am J Physiol. 1999;277:H351–62.
Neumann E, Kakorin S, Toensing K. Fundamentals of electroporative delivery of drugs and genes. Bioelectrochem Bioenerg. 1999;48:3–16.
Teissié J, Eynard N, Gabriel B, et al. Electropermeabilization of cell membranes. Adv Drug Deliv Rev. 1999;35:3–19.
Burnett P, Robertson JK, Palmer JM, et al. Fluorescence imaging of electrically stimulated cells. J Biomol Screen. 2003;8:660–7.
Sharma V, Tung L. Ionic currents involved in shock-induced nonlinear changes in transmembrane potential responses of single cardiac cells. Pflugers Arch. 2004;449:248–56.
Huang CJ, Harootunian A, Maher MP, et al. Characterization of voltage-gated sodium-channel blockers by electrical stimulation and fluorescence detection of membrane potential. Nat Biotechnol. 2006;24:439–46.
Pauly H, Schwan HP. Über die Impedanz einer Suspension von kugelformigen Teilchen mit einer Schale. Z Naturforsch B. 1959;14:125–31.
Kotnik T, Bobanović F, Miklavčič D. Sensitivity of transmembrane voltage induced by applied electric fields – a theoretical analysis. Bioelectrochem Bioenerg. 1997;43:285–91.
Hibino M, Shigemori M, Itoh H, et al. Membrane conductance of an electroporated cell analyzed by submicrosecond imaging of transmembrane potential. Biophys J. 1991;59:209–20.
Hibino M, Itoh H, Kinosita Jr K. Time courses of cell electroporation as revealed by submicrosecond imaging of transmembrane potential. Biophys J. 1993;64:1789–800.
Tekle E, Astumian RD, Chock PB. Selective and asymmetric molecular-transport across electroporated cell-membranes. Proc Natl Acad Sci USA. 1994;91:11512–6.
Gabriel B, Teissié J. Direct observation in the millisecond time range of fluorescent molecule asymmetrical interaction with the electropermeabilized cell membrane. Biophys J. 1997;73:2630–7.
Gabriel B, Teissié J. Time courses of mammalian cell electropermeabilization observed by millisecond imaging of membrane property changes during the pulse. Biophys J. 1999;76:2158–65.
Schwan HP. Electrical properties of tissue and cell suspensions. Adv Biol Med Phys. 1957;5:147–209.
Grosse C, Schwan HP. Cellular membrane potentials induced by alternating fields. Biophys J. 1992;63:1632–42.
Kotnik T, Miklavčič D, Slivnik T. Time course of transmembrane voltage induced by time-varying electric fields – a method for theoretical analysis and its application. Bioelectrochem Bioenerg. 1998;45:3–16.
Kotnik T, Miklavčič D. Second-order model of membrane electric field induced by alternating external electric fields. IEEE Trans Biomed Eng. 2000;47:1074–81.
Fluhler E, Burnham VG, Loew LM. Spectra, membrane binding, and potentiometric responses of new charge shift probes. Biochemistry. 1985;24:5749–55.
Gross D, Loew LM, Webb W. Optical imaging of cell membrane potential changes induced by applied electric fields. Biophys J. 1986;50:339–48.
Loew LM. Voltage sensitive dyes: measurement of membrane potentials induced by DC and AC electric fields. Bioelectromagnetics. 1992;Suppl 1:179–89.
Pucihar G, Kotnik T, Miklavčič D. Measuring the induced membrane voltage with di-8-ANEPPS. J Visual Exp 2009;33:1659.
Bernhard J, Pauly H. Generation of potential differences across membranes of ellipsoidal cells in an alternating electrical field. Biophysik. 1973;10:89–98.
Kotnik T, Miklavčič D. Analytical description of transmembrane voltage induced by electric fields on spheroidal cells. Biophys J. 2000;79:670–9.
Gimsa J, Wachner D. Analytical description of the transmembrane voltage induced on arbitrarily oriented ellipsoidal and cylindrical cells. Biophys J. 2001;81:1888–96.
Fear EC, Stuchly MA. Modeling assemblies of biological cells exposed to electric fields. IEEE Trans Biomed Eng. 1998;45:1259–71.
Buitenweg JR, Rutten WL, Marani E. Geometry-based finite-element modeling of the electrical contact between a cultured neuron and a microelectrode. IEEE Trans Biomed Eng. 2003;50:501–9.
Valič B, Golzio M, Pavlin M, et al. Effect of electric field induced transmembrane potential on spheroidal cells: theory and experiment. Eur Biophys J. 2003;32:519–28.
Pucihar G, Kotnik T, Valič B, et al. Numerical determination of transmembrane voltage induced on irregularly shaped cells. Ann Biomed Eng. 2006;34:642–52.
Pucihar G, Miklavčič D, Kotnik T. A time-dependent numerical model of transmembrane voltage inducement and electroporation of irregularly shaped cells. IEEE Trans Biomed Eng. 2009;56:1491–501.
Teruel MN, Meyer T. Electroporation-induced formation of individual calcium entry sites in the cell body and processes of adherent cells. Biophys J. 1997;73:1785–96.
Rols MP, Teissié J. Electropermeabilization of mammalian cells. Quantitative analysis of the phenomenon. Biophys J. 1990;58:1089–98.
Wolf H, Rols MP, Boldt E, et al. Control by pulse duration of electric-field mediated gene transfer in mammalian cells. Biophys J. 1994;66:524–31.
Rols MP, Teissié J. Electropermeabilization of mammalian cells to macromolecules: control by pulse duration. Biophys J. 1998;75:1415–23.
Canatella PJ, Karr JF, Petros JA, et al. Quantitative study of electroporation-mediated molecular uptake and cell viability. Biophys J. 2001;80:755–64.
Maček-Lebar A, Miklavčič D. Cell electropermeabilization to small molecules in vitro: control by pulse parameters. Radiol Oncol. 2001;35:193–202.
Čemažar M, Jarm T, Miklavčič D, et al. Effect of electric-field intensity on electropermeabilization and electrosensitivity of various tumor-cell lines in vitro. Electro Magnetobiol. 1998;17:261–70.
Troiano GC, Tung L, Sharma V, et al. The reduction in electroporation voltages by the addition of a surfactant to planar lipid bilayers. Biophys J. 1998;75:880–8.
Kandušer M, Fošnarič M, Šentjurc M, et al. Effect of surfactant polyoxyethylene glycol (C12E8) on electroporation of cell line DC3F. Colloid Surface A. 2003;214:205–17.
Sukharev SI, Klenchin VA, Serov SM, et al. Electroporation and electrophoretic DNA transfer into cells. Biophys J. 1992;63:1320–7.
Satkauskas S, André F, Bureau MF, et al. Electrophoretic component of electric pulses determines the efficacy of in vivo DNA electrotransfer. Human Gene Ther. 2005;16:1194–201.
Kandušer M, Miklavčič D, Pavlin M. Mechanisms involved in gene electrotransfer using high- and low-voltage pulses – an in vitro study. Bioelectrochemistry. 2009;74:265–71.
Tekle E, Astumian RD, Chock PB. Electroporation by using bipolar oscillating electric field: an improved method for DNA transfection of NIH 3T3 cells. Proc Natl Acad Sci USA. 1994;88:4230–4.
Kotnik T, Mir LM, Flisar K, et al. Cell membrane electropermeabilization by symmetrical bipolar rectangular pulses. Part I. Increased efficiency of permabilization. Bioelectro-chemistry. 2001;54:83–90.
Kotnik T, Miklavčič D, Mir LM. Cell membrane electropermeabilization by symmetrical bipolar rectangular pulses: Part II. Reduced electrolytic contamination. Bioelectrochemistry. 2001;54:91–5.
Chang DC. Cell poration and cell fusion using an oscillating electric field. Biophys J. 1989;56:641–52.
Chang DC, Gao PQ, Maxwell BL. High efficiency gene transfection by electroporation using a radio-frequency electric field. Biochim Biophys Acta. 1991;1092:153–60.
Xie TD, Tsong TY. Study of mechanisms of electric field-induced DNA transfection. II. Transfection by low-amplitude, low-frequency alternating electric fields. Biophys J. 1990;58:897–903.
Kotnik T, Pucihar G, Reberšek M, et al. Role of pulse shape in cell membrane electropermeabilization. Biochim Biophys Acta. 2003;1614:193–200.
Pucihar G, Mir LM, Miklavčič D. The effect of pulse repetition frequency on the uptake into electropermeabilized cells in vitro with possible applications in electrochemotherapy. Bioelectrochemistry. 2002;57:167–72.
Pucihar G, Kotnik T, Kandušer M, et al. The influence of medium conductivity on electropermeabilization and survival of cells in vitro. Bioelectrochemistry. 2001;54:107–15.
Nicotera C, Bellomo G, Orrenius S. Calcium-mediated mechanisms in chemically induced cell death. Annu Rev Pharmacol Toxicol. 1992;32:449–70.
Golzio M, Mora MP, Raynaud C, et al. Control by osmotic pressure of voltage-induced permeabilization and gene transfer in mammalian cells. Biophys J. 1998;74:3015–22.
Rols MP, Delteil C, Serin G, et al. Temperature effects on electrotransfection of mammalian cells. Nucleic Acids Res. 1994;22:540.
Susil R, Šemrov D, Miklavčič D. Electric field induced transmembrane potential depends on cell density and organization. Electro Magnetobiol. 1998;17:391–9.
Pavlin M, Pavšelj N, Miklavčič D. Dependence of induced transmembrane potential on cell density, arrangement, and cell position inside a cell system. IEEE Trans Biomed Eng. 2002;49:605–12.
Pucihar G, Kotnik T, Teissié J, et al. Electroporation of dense cell suspensions. Eur Biophys J. 2007;36:173–85.
Reberšek M, Faurie C, Kandušer M, et al. Electroporator with automatic change of electric field direction improves gene electrotransfer in vitro. Biomed Eng Online. 2007;6(25):1–11.
Trontelj K, Reberšek M, Kandušer M, et al. Optimization of bulk cell electrofusion in vitro for production of human–mouse heterohybridoma cells. Bioelectrochemistry. 2008;74:124–9.
Towhidi L, Kotnik T, Pucihar G, et al. Variability of the minimal transmembrane voltage resulting in detectable membrane electroporation. Electromagn Biol Med. 2008;27:372–85.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2011 Springer Science+Business Media, LLC
About this chapter
Cite this chapter
Kotnik, T., Pucihar, G., Miklavčič, D. (2011). The Cell in the Electric Field. In: Kee, S., Gehl, J., Lee, E. (eds) Clinical Aspects of Electroporation. Springer, New York, NY. https://doi.org/10.1007/978-1-4419-8363-3_3
Download citation
DOI: https://doi.org/10.1007/978-1-4419-8363-3_3
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-4419-8362-6
Online ISBN: 978-1-4419-8363-3
eBook Packages: MedicineMedicine (R0)