Electric field induced transient pores in phospholipid bilayer vesicles

Teissié, Justin; Tsong, Tian Yow

doi:10.1021/bi00509a022

Cited by 303 publications

(167 citation statements)

References 24 publications

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“…This potential is induced within 1 pus on the vesicle (19) (20) and Azzone et aL (21). The absence of synthesis with a short electric pulse (12 Us) provides an additional feature for the model.…”

Section: Resultsmentioning

confidence: 99%

Synthesis of adenosine triphosphate in respiration-inhibited submitochondrial particles induced by microsecond electric pulses

Teissié

Knox

Tsong

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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Phosphorylation of ADP to ATP was induced in nonrespiring submitochondrial particles (SMP) from rat liver by the application of electric pulses with field strengths of kV/ cm and a decay time of60 pas. In all cases respiration was inhibited completely by using cyanide or rotenone. Newly formed ATP was measured by two independent methods, (i) the luciferase/luciferin bioluminescence assay and (ii) synthesis of [3P]ATP from ADP and 32Pi. Both The mechanism by which energy released during respiration is used to synthesize the terminal phosphate bond of ATP has been the subject ofinvestigation for many years. Recent studies have focused on the role of the mitochondrial inner membrane in the energy transducing process (1). In addition to organizing the complex multienzyme systems of electron transport, such a membrane has the ability to store energy, in the form ofchemical concentration gradients and ofcharge separations. The formation ofan electrochemical gradient and the subsequent conversion of its energy into the chemical bond energy ofATP has been postulated as the basic mechanism ofenergy transduction (1).Experiments have been designed to test the above hypothesis. Jagendorf and Uribe (2) were able to form a pH gradient (ApH) across the chloroplast thylakoid membrane. By rapidly transferring chloroplasts from an acidic to a basic medium in the dark they induced ATP synthesis. Similar experiments were performed on submitochondrial particles (SMP) from beef heart, using a proton and K+ concentration jump (3). In fact, reasonable rates of ATP synthesis were achieved by a K+ concentration jump alone, but the presence ofa K+ ionophore was found to be essential. In these experiments effects of changes in proton or ion concentrations on the membrane enzyme activity were difficult to assess. Artificial ionophores may also alter membrane structure.An alternative approach is to use high-voltage electric pulse techniques (see, for example, ref. 4). By exposing a suspension of cells or organelles to an electric field it is possible to directly impose an electrical potential difference across the membrane (Alf). This A4i is position dependent, and different locales at the membrane surface may experience different levels of Afi (see Fig. 1 and Discussion). Witt and his coworkers (5) have employed this technique in the chloroplast system and observed that a 30-ms electric pulse with a field strength of 1.1 kV/cm induced ATP synthesis in the dark. Their results demonstrate that externally applied electrical energy may be utilized for ATP synthesis by the light energy transducing system. Experiments with SMP are inherently more difficult because oftheir smaller size. Ten to 30 times greater field strength must be used to generate the same A+. Joule heating thus becomes more severe. In this communication, we report the induction ofATP synthesis in SMP (from rat liver) by a microsecond electric pulse, in the presence ofcyanide or rotenone to completely inhibit respiration. By using two independent and complementary me...

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Section: Resultsmentioning

confidence: 99%

Synthesis of adenosine triphosphate in respiration-inhibited submitochondrial particles induced by microsecond electric pulses

Teissié

Knox

Tsong

et al. 1981

Proc. Natl. Acad. Sci. U.S.A.

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“…This suggests that transfection depends on the creation of pores between the cell and specifically bound liposomes permitting transfer of locally high concentrations of plasmid DNA. Electroporation is known to form pores in both plasma membranes (24) and lipid vesicles (50). Alternatively, transfection can be the result of direct fusion of the cell and the bound liposome, since electric fields can also induce fusion of lipid membranes (50)(51)(52)(53)(54).…”

Section: Discussionmentioning

confidence: 99%

Gene transfer from targeted liposomes to specific lymphoid cells by electroporation.

Machy¹,

Lewis²,

McMillan³

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

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Large unilamellar liposomes, coated with protein A and encapsulating the gene that confers resistance to mycophenolic acid, were used as a model system to demonstrate gene transfer into specific lymphoid cells. Protein A, which selectively recognizes mouse IgG2a antibodies, was coupled to liposomes to target them specifically to defined cell types coated with IgG2a antibody. Protein A-coated liposomes bound human B lymphoblastoid cells preincubated with a mouse IgG2a anti-HLA monoclonal antibody but failed to adhere to cells challenged with an irrelevant (anti-H-2) antibody of the same isotype or to cells incubated in the absence of antibody. Transfection of target cells bound to protein A-coated liposomes was achieved by electroporation. This step was essential since only electroporated cells survived in a selective medium containing mycophenolic acid. Transfection efficiency with electroporation and targeted liposomes was as efficient as conventional procedures that used unencapsulated plasmids free in solution but, in the latter case, cell selectivity is not possible. This technique provides a methodology for introducing defmed biological macromolecules into specific cell types.Transfection ofeukaryotic cells by DNA is a useful technique to increase our understanding of gene expression and regulation and of the function of given molecules in particular cell types. Early transfection studies relied upon two procedures to introduce exogenous genetic material into cells, (i) natural transfection, accomplished by infecting cells with genetically manipulated but intact virus (1, 2) and (ii) artificial procedures involving temporary physical or chemical perturbations of plasma membranes to permit entry of DNA. The second technique that uses complexes of DNA with calcium phosphate (3), DEAE-dextran (4, 5), or polyornithine (4) (24,25), or tungsten microprojectiles (26). Each of these procedures is distinguished by its own spectrum of advantages and disadvantages, including efficiency, toxicity, technical difficulty, equipment needs, and specificity.In this study we show that lymphocytes can be specifically transfected in vitro by using an electric field (electroporation) and targeted liposomes containing DNA. As a model system we used large unilamellar liposomes in which plasmid DNA carrying the bacterial xanthine guanine phosphoribosyltransferase (XGPRT) gene was encapsulated. These liposomes were directed to target cells by monoclonal antibodies and specific transfection was achieved by electroporation. The technique is simple and specific and can be used to introduce genetic material and macromolecules into other cell types. MATERIALS AND METHODSCells. The human Burkitt lymphoma cell line BJAB was cultured in Dulbecco's modified Eagle medium (DMEM, GIBCO) supplemented with 5% (vol/vol) heat-inactivated fetal calf serum, 2 mM glutamine, and gentamicin at 37°C in an atmosphere of 7% C02/93% air.Monoclonal Antibodies. B1.23.2 and H100-5/28 are monoclonal IgG2a mouse antibodies. B1.23.2 is directed against no...

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“…the sum between the resting value of cell membrane ΔΨo and the electroinduced value ΔΨi) reaches threshold values close to 250 mV, membranes become permeable. [25][26] Permeabilization is controlled by the field strength. Field intensity larger than a critical value (Ep) must be applied to the cell suspension.…”

Section: Critical Parameters Affecting Electropermeabilizationmentioning

confidence: 99%

Time dependence of electric field effects on cell membranes. A review for a critical selection of pulse duration for therapeutical applications

Teissié¹,

Escoffre²,

Rols³

et al. 2008

Radiology and Oncology

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Background. Electropulsation is one of the non-viral methods successfully used to transfer drugs and genes into living cells in vitro as in vivo. This approach shows promise in field of gene and cellular therapies. This presentation first describes the temporal factors controlling electropermeabilization to small molecules (< 4kDa) and then the processes supporting DNA transfer in vitro. The description of in vitro events brings our attention on the processes occurring before (s), during (ms) and after electropulsation (ms to hours) ofhas the main advantages of being easy to use, fast, reproducible and safe.While during 30 years due to technological limits, pulse duration was always larger than 1 microsecond, the recent availability of high voltage (tens of kV) nanosecond long pulse generators opens the way to a new approach. Very fast perturbations under strong fields are induced in the membrane organization. 16,17 A new field of development is now present for electropermeabilization and promising results for clinical applications were reported.One of the limiting problems remains that very few is known on the physicochemical mechanisms supporting the reorganisation of the cell membrane. The molecular target of the field effect remains unclear.The present review focuses on the critical role played by the pulse duration in the electropermeabilization to small molecules (< 4kDa) and on its support to the processes associated to DNA transfer in vitro. Pulse durations are easy to adjust for an optimization of the clinical target: electrochemotherapy, irreversible electropermeabilization or gene therapy as suggested as a final conclusion. Electropermeabilization Theory of membrane potential difference modulation.An external electric field modulates the membrane potential difference. 18 From the physical point of view, a cell can be described as a spherical capacitor which is charged by the external electrical field. The transmembrane potential difference induced by the electric field, ΔΨi is a complex function g(λ) of the specific conductivities of the membrane (λ m ), the pulsing buffer (λ out ) and the cytoplasm (λ cyt ), the membrane thickness and the cell size. Thus, 1 ΔΨi = f. g (λ). r. E.cosθ [1] in which θ designates the angle between the direction of the normal to the membrane at the considered point on the cell surface and the field direction, E the field intensity, r the radius of the cell and f, which is a shape factor (a cell being a spheroid). Therefore, ΔΨi is not uniform on the cell surface. It is maximum at the positions of the cell facing the electrodes. These physical predictions were checked experimentally by videomicroscopy by using the potential difference sensitive fluorescent probes. [19][20][21] The pulse duration plays a critical role when shorter than the capacitive loading time of the membrane. In the previous part of the paper, it was considered that the pulse was long enough to bring the potential steady state value. The loading time τ load brings a limit in this description. 1 ΔΨi = f. ...

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Electric field induced transient pores in phospholipid bilayer vesicles

Cited by 303 publications

References 24 publications

Synthesis of adenosine triphosphate in respiration-inhibited submitochondrial particles induced by microsecond electric pulses

Synthesis of adenosine triphosphate in respiration-inhibited submitochondrial particles induced by microsecond electric pulses

Gene transfer from targeted liposomes to specific lymphoid cells by electroporation.

Time dependence of electric field effects on cell membranes. A review for a critical selection of pulse duration for therapeutical applications

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