Membrane Electrical Parameters Research Articles

Interaction of LL-37 human cathelicidin peptide with a model microbial-like lipid membrane

The only representative of cathelicidin peptides in humans is LL-37, a multifunctional antimicrobial peptide (AMP) that is a part of the innate immune response. Details of the LL-37 direct activity against pathogens are not well understood at the molecular level. Here, we present research on the mechanism of interaction between LL-37 and a model multicomponent bilayer lipid membrane (BLM), mimicking microbial cell membrane. Electrochemical impedance spectroscopy (EIS), high-resolution atomic force microscopy (AFM) imaging, and polarization-modulation infrared reflection–absorption spectroscopy (PM-IRRAS) were applied to study the peptide influence on a model microbial-like membrane. We show that LL-37 causes changes in the phospholipid molecules conformation and orientation, leading to membrane disintegration, significantly affecting the membrane electrical parameters, such as capacitance and resistance. High-resolution AFM imaging shows topographical and mechanical effects of such disintegration, while PM-IRRAS data indicates that introduction of LL-37 causes changes in the phospholipid acyl chains from all-trans to gauche conformations. Moreover, the presence of LL-37 significantly alters the value of the phospholipid tilt angle. Altogether, our results suggest a “carpet” membrane dissolution followed by a detergent-like membrane disruption mechanism upon LL-37 activity. This research gives a novel insight into the understanding of LL-37 influence on multicomponent model membranes and a promising contribution to the development of LL-37-derived therapeutic agents against drug-resistant bacteria.

Plasticization-induced oriented micro-channels within polymer inclusion membranes for facilitating Cu(II) transport

Although polymer inclusion membranes (PIMs) have been potentially used for facilitating the ion transport, the internal structure evolution of the PIMs is still controversial. This work focused on the effect of the composition and structure of PIMs on the transport of Cu(II), which were composed of poly(vinyl chloride) (PVC) as skeleton, 2-nitrophenyloctyl ether (NPOE) as the plasticizer, and commercial LIX84I as the carrier. The results suggested that the transport flux of Cu(II) is significantly dependent on the membranous composition. While correlating the membranous composition, the microstructural information of the PVC-based PIMs was explored and evaluated using X-ray diffraction (XRD), scanning electron microscopy (SEM), small-angle X-ray scattering (SAXS), and electrochemical impedance spectroscopy (EIS), respectively. XRD patterns and SAXS spectral features indicated that the significant structure transition occurred while increasing the NPOE content. Micro-channels of around 9 nm average diameter were clearly disclosed by SAXS for the PIM containing 40 wt% PVC, 30 wt% NOPE and 30 wt% LIX84I. And the well-organized pathway within in the PIMs was essential for efficient Cu(II) transport. The membrane's electrical parameters including dielectric constant and conductivity were also found to be depended on the membranous composition and structure. Besides, the selective transport of Cu(II) towards Ni(II) and Co(II) with PIM were further examined.

Tuning physicochemical, electrochemical and transport characteristics of polymer inclusion membrane by varying the counter-anion of the ionic liquid Aliquat 336

Polymer inclusion membranes (PIMs) have been prepared using cellulose triacetate (CTA) as polymer and derivatives of the commercial ionic liquid (IL) Aliquat 336, trioctyl methylammonium chloride (AlqCl), as extractants. The different ILs were prepared by exchanging chloride anion from AlqCl for other more lipophilic anions, obtaining IL derivatives AlqNO3 and AlqSCN. PIMs containing these extractants at two different concentrations (30% and 60%) were prepared and characterized using X-ray photoelectron spectroscopy (XPS), infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), contact angle and impedance spectroscopy to obtain information on the material properties of both the surface and bulk membranes. The IL counter-anion affects the membrane's electrical parameters (dielectric constant and conductivity) as well as its hydrophobic character, which also depends on the IL concentration in the PIM formulation. Passive transport across the different PIMs using HCl and NaCl aqueous solutions was also considered, and the results obtained seem to be directly related to the hydrophilic/hydrophobic character of the studied membranes.

Physicochemical modelling of the surface-active phospholipid bilayer relative to acid-base equilibria

The objective of this research was to evaluate interactions of hydrogen and hydroxide ions with bilayer membranes formed from phosphatidylcholine and lysophosphatidylcholine. Theoretical deliberations were successfully verified by experimental research studies that were conducted using electrochemical impedance spectroscopy and chronopotentiometry methods. Electrical capacitance and electrical resistance measurements were made on the membranes in various pH electrolyte solutions. The theoretical calculations were based on previously derived two theoretical models. In both models the contributions of the individual lipid molecule forms to the membrane electrical parameters were assumed to be additive.

Accelerating bioelectric functional development of neural stem cells by graphene coupling: Implications for neural interfacing with conductive materials

In order to govern cell-specific behaviors in tissue engineering for neural repair and regeneration, a better understanding of material-cell interactions, especially the bioelectric functions, is extremely important. Graphene has been reported to be a potential candidate for use as a scaffold and neural interfacing material. However, the bioelectric evolvement of cell membranes on these conductive graphene substrates remains largely uninvestigated. In this study, we used a neural stem cell (NSC) model to explore the possible changes in membrane bioelectric properties - including resting membrane potentials and action potentials - and cell behaviors on graphene films under both proliferation and differentiation conditions. We used a combination of single-cell electrophysiological recordings and traditional cell biology techniques. Graphene did not affect the basic membrane electrical parameters (capacitance and input resistance), but resting membrane potentials of cells on graphene substrates were more strongly negative under both proliferation and differentiation conditions. Also, NSCs and their progeny on graphene substrates exhibited increased firing of action potentials during development compared to controls. However, graphene only slightly affected the electric characterizations of mature NSC progeny. The modulation of passive and active bioelectric properties on the graphene substrate was accompanied by enhanced NSC differentiation. Furthermore, spine density, synapse proteins expressions and synaptic activity were all increased in graphene group. Modeling of the electric field on conductive graphene substrates suggests that the electric field produced by the electronegative cell membrane is much higher on graphene substrates than that on control, and this might explain the observed changes of bioelectric development by graphene coupling. Our results indicate that graphene is able to accelerate NSC maturation during development, especially with regard to bioelectric evolvement. Our findings provide a fundamental understanding of the role of conductive materials in tuning the membrane bioelectric properties in a graphene model and pave the way for future studies on the development of methods and materials for manipulating membrane properties in a controllable way for NSC-based therapies.

Polarizability of red blood cells with an anisotropic membrane

We predict the complex polarizability of a realistic model of a red blood cell (RBC), with an inhomogeneous dispersive and anisotropic membrane. In this model, the frequency-dependent complex electrical parameters of the individual cell layers are described by the Debye equation while the dielectric anisotropy of the cell membrane is taken into account by the different permittivities along directions normal and tangential to the membrane surface. The realistic shape of the RBC is described in terms of the Jacobi elliptic functions. To calculate the polarizability, we evoke the effective dipole moment method to determine the cell internal electric field distribution, employing an adaptive finite-element numerical approach. We have furthermore investigated the influence of the anisotropic membrane and dispersive electrical parameters of each individual cell layer on the total complex polarizability. Our findings suggest that the individual layer contribution depends on two factors: the volume of the layer and the associated induced electric field, which in turn is influenced by other layers of the cell. These results further show that the average polarizability spectra of the cell are significantly impacted by the anisotropy and associated dispersion of the cellular compartments.

Electrochemical Characterization of an Asymmetric Nanofiltration Membrane with NaCl and KCl Solutions: Influence of Membrane Asymmetry on Transport Parameters

Electrochemical characterization of a nanofiltration asymmetric membrane was carried out by measuring membrane potential, salt diffusion, and electrical parameters (membrane electrical resistance and capacitance) with the membrane in contact with NaCl and KCl solutions at different concentrations (10−3≤c(M)≤5×10−2). From these experiments characteristic parameters such as the effective concentration of charge in the membrane, ionic transport numbers, and salt and ionic permeabilities across the membrane were determined. Membrane electrical resistance and capacitance were obtained from impedance spectroscopy (IS) measurements by using equivalent circuits as models. This technique allows the determination of the electrical contribution associated with each sublayer; then, assuming that the dense sublayer behaves as a plane capacitor, its thickness can be estimated from the capacitance value. The influence of membrane asymmetry on transport parameters have been studied by carrying out measurements for the two opposite external conditions. Results show that membrane asymmetry strongly affects membrane potential, which is attributed to the Donnan exclusion when the solutions in contact with the dense layer have concentrations lower than the membrane fixed charge (Xef≈−0.004 M), but for the reversal experimental condition (high concentration in contact with the membrane dense sublayer) the membrane potential is practically similar to the solution diffusion potential. The comparison of results obtained for both electrolytes agrees with the higher conductivity of KCl solutions. On the other hand, the influence of diffusion layers at the membrane/solution interfaces in salt permeation was also studied by measuring salt diffusion at a given NaCl concentration gradient but at five different solutions stirring rates.

Translocation of polysialic acid across model membranes: kinetic analysis and dynamic studies.

Transmembrane translocation of polyion homopolymers takes place in the case of polyanionic polysialic acid (polySia), polyanionic polynucleotides and polycationic polypeptides. The purpose of this work was to determine the role of membrane electrical parameters on the kinetics of polyion translocation, the influence of polysialic acid on ion adsorption on positively charged membrane surface and the dynamics of the phospholipid hydrocarbon chains and choline group by using 1H-NMR. The analysis of polyion translocation was performed by using the electrical equivalent circuit of the membrane for the initial membrane potential equal to zero. The changes in polysialic acid flux was up to 75% after 1 ms in comparison with the zero-time flux. Both a decrease of membrane conductance and an increase of polyion chain length resulted in the diminution of this effect. An increase of praseodymium ions adsorption to positively charged liposomes and an increase of the rate of segmental movement of the -CH2 and -CH3 groups, and the choline headgrup of lipid molecules, was observed in the presence of polySia. The results show that the direction of the vectorial polyion translocation depends both on the membrane electrical properties and the degree of polymerization of the polymer, and that polysialic acid can modulate the degree of ion adsorption and the dynamics of membrane lipids.

Polylysine induces changes in membrane electrical properties of K562 cells.

The ionic environment of the cell membrane is of extreme importance in maintaining cell integrity and the numerous functions necessary for cell growth, differentiation, etc., as well as in cell-biomaterial interactions. In this study, the effects of polylysine (a basic poly-amino acid with a net positive charge which is often used to coat biomaterials surfaces) on the erythroleukemic K562 cell membrane were investigated. In particular, the effects of this polycation were evaluated using dielectric relaxation studies in the radiofrequency range with which it is possible to measure both active ionic transport across the cell membrane (membrane conductivity) and the static charge distribution present on the cell surface due to the structural components of the cell membrane (membrane permittivity). The conductivity of the cytosol can also be determined. The results demonstrate that while the conductivity of the cytosol is not significantly altered, both the conductivity and permittivity of the K562 cell membrane are varied by exposure of these cells to polylysine. These observations indicate that both active ionic transport and the type, quantity, or distribution of membrane components such as lipids, proteins, and polysaccharides may also be altered. Although the precise mechanisms by which these variations in K562 cells occur are unknown, it can be hypothesized that changes in the growth characteristics of these cells may be in part responsible. In particular, as demonstrated by light microscopic examination of K562 cells directly in the culture flasks, the cells in polylysine-coated flasks do not grow in suspension as do the controls, but rather show anchorage-dependent-like behavior. It is this important change from suspension to monolayer growth induced by polylysine that may be responsible for the changes in membrane electrical parameters.

Changes in membrane electrical parameters of yeast following chemical treatment for protoplast isolation

The yeast strains Saccharomyces species and Candida tropicalis were used. In each case, the electrorotation spectra were recorded for normal untreated yeast cells, β-mercaptoethanol-treated cells and isolated protoplasts. In order to obtain realistic information on the passive electrical membrane parameters associated with each step of protoplast isolation, a theoretical analysis of the simulated electrorotation spectra for shelled ellipsoids was performed. As a consequence, the parameters chosen for fitting of the experimental spectra were the electrical conductivities of the cell compartments, the membrane permittivity and the scaling factor. β-Mercaptoethanol was found to have significant action at the membrane level: (1) at high concentration (2%), σ m was increased dramatically; (2) in mild incubation conditions (0.1%), σ m was decreased slightly. Treatment with the enzyme complex led to a decrease in internal conductivity, probably due to a loss of the ionic equilibrium balance.

A static magnetic field does not affect the dielectric properties of chick embryo myoblast membranes.

We have recently demonstrated using dielectic relaxation studies in the radiowave frequency range that sinusoidal 50 Hz magnetic fields (with intensities ranging from 1 to 10 mT) induce a nonlinear change in both membrane conductivity and permittivity of primary chick embryo myoblasts in vitro. It was the aim of the present study to determine if a DC-induced static magnetic field is capable of generating similar variations in the membrane conductivity and/or the membrane permittivity of chick embryo myoblasts. The results indicate that when the myogenic cells are exposed to a static magnetic field of either 1, 3 or 5 mT (values comparable with the previous extremely low frequency study), no changes in the membrane electrical parameters can be observed with respect to controls. Differences in the characteristics of static and extremely low frequency fields as well as the possible mechanisms underlying the contrasting results with these two types of magnetic fields are discussed.

Block of GABA-transaminase modifies GABAergic transmission at the crayfish synapses.

1. The cytosolic concentration of a neurotransmitter is believed to be an important factor determining its release. The effects of ethanolamine-O-sulfate (EOS), a gamma-aminobutyric acid (GABA)-transaminase blocker, on GABAergic postsynaptic and presynaptic inhibitory neurotransmission were examined in the crayfish opener neuromuscular synapses. 2. Intracellular recordings of evoked excitatory (EPSPs) and inhibitory postsynaptic potentials (IPSPs) as well as loose macropatch clamp measurements of excitatory (EPSCs) and inhibitory postsynaptic currents (IPSCs) were used to evaluate the effects of the drug, which was applied exclusively to the nerve bundle. 3. Under normal conditions, a stimulus train to the inhibitor before the excitor stimulation elicited two phases of inhibition: 1) a large reduction in EPSP amplitude associated with a decrease in its time constant of decay (tau D) at time intervals of 0-15 ms and 2) a moderate decrease in EPSP amplitude with a small change in EPSP tau D at intervals of 15-90 ms. EOS treatment selectively increased the inhibition of phase 2. 4. The muscle membrane electrical parameters and the existing postsynaptic tonic release of GABA were not affected by the drug. 5. EOS did not alter the IPSP's parameters such as amplitude, reversal potential, and conductance. 6. Quantal analysis of single IPSCs revealed no significant changes in the statistical parameters such as quantum size (q), quantal content (m), number of active zones (n), or probability of release (p). 7. Quantal analysis of EPSCs, released after interaction with the inhibitor, did exhibit a large reduction in m without any effect on q. 8. These results demonstrate that EOS has a specific and differential effect on neural transmission in two synapses of the same axon: it increases presynaptic inhibition without significant effect on the postsynaptic inhibitory mechanism.

Non-linear dependence of the dielectric properties of chick embryo myoblast membranes exposed to a sinusoidal 50 Hz magnetic field

Primary chick embryo myoblasts can be a useful tool for studying the developmental events which accompany myoblast differentiation, particularly myoblast membrane fusion. To determine whether the electrical properties and/or fusion in these systems are affected by 50 Hz magnetic fields, chick embryo myoblast cultures were exposed to B-field intensities ranging from 1 to 10 mT. The electrical parameters of the myoblasts, i.e. membrane conductivity, membrane permittivity and the conductivity of the cell interior (cytosol) were determined by the analysis of conductivity dispersion data in the radio frequency range (10 kHz-100 MHz). Preliminary results indicate that the time of fusion (60 h) is not affected by these fields, but that the absolute values of the two membrane electrical parameters are affected. In particular, a B-field intensity-dependent decrease was observed. The maximum effect resulted after a 1 h exposure to a magnetic flux density of about 5 mT. The conductivity of the cytosol remained unchanged. These data seem to indicate that exposure to 50 Hz magnetic fields affects both static and dynamic membrane properties in primary chick embryo myoblasts.

Dielectrophoresis and electrorotation of neurospora slime and murine myeloma cells

Dielectrophoresis and electrorotation are commonly used to measure dielectric properties and membrane electrical parameters of biological cells. We have derived quantitative relationships for several critical points, defined in Fig. A 1, which characterize the dielectrophoretic spectrum and the electrorotational spectrum of a cell, based on the single-shell model (Pauly, H., and H.P. Schwan, 1959. Z. Naturforsch. 14b:125-131; Sauer, F.A. 1985. Interactions between Electromagnetic Field and Cells. A. Chiabrera, C. Nicolini, and H.P. Schwan, editors. Plenum Publishing Corp., New York. 181-202). To test these equations and to obtain membrane electrical parameters, a technique which allowed simultaneous measurements of the dielectrophoresis and the electrorotation of single cells in the same chamber, was developed and applied to the study of Neurospora slime and the Myeloma Tib9 cell line. Membrane electrical parameters were determined by the dependence of the first critical frequency of dielectrophoresis (fct1) and the first characteristic frequency of electrorotation (fc1) on the conductivity of the suspending medium. Membrane conductances of Neurospora slime and Myeloma also were found to be 500 and 380 S m-2, respectively. Several observations indicate that these cells are more complex than that described by the single-shell model. First, the membrane capacities from fct1 (0.81 x 10(-2) and 1.55 x 10(-2) F m-2 for neurospora slime and Myeloma, respectively) were at least twice those derived from fc1. Second, the electrorotation spectrum of Myeloma cells deviated from the single-shell like behavior. These discrepancies could be eliminated by adapting a three-shell model (Furhr, G., J. Gimsa, and R. Glaser. 1985. Stud. Biophys. 108:149-164). Apparently, there was more than one membrane relaxation process which could influence the lower frequency region of the beta-dispersion. fct1 of Myeloma in a medium of given external conductivity were found to be similar for most cells, but for some a dramatically increased fct1 was recorded. Model analysis suggested that a decrease in the cytoplasmatic conductivity due to a drastic ion loss in a cell could cause this increase in fct1. Model analysis also suggested that the electrorotation spectrum in the counter-field rotation range and fc1 would be more sensitive to conductivity changes of the cytoplasmic fluid and to the influence of internal membranes than would fct1, although the latter would be sensitive to changes in capacitance of the cytoplasmic membranes.

Modulation of rat skeletal muscle chloride channels by activators and inhibitors of protein kinase C.

The membrane electrical parameters and component conductances of rat extensor digitorum longus muscle fibres were studied in vitro at 30 degrees C with standard two microelectrode square pulse cable analysis in the presence of protein kinase C (PKC) activators and inhibitors. The PKC activator, 4-beta-phorbol-12,13 dibutyrate (4-beta-PDB), (2-90 nM) blocked up to 67% chloride conductance (GCl) in rat skeletal muscle fibres and induced myotonic hyperexcitability. The concentration necessary to produce a 50% block of the membrane GCl was 23 nM. The "inactive" 4-alpha-phorbol-12,13 dibutyrate had no effect at 2 microM. The blocking effect of 4-beta-PDB on GCl was prevented by preincubation of the preparations with the PKC inhibitors, staurosporine (1-5 microM) and tetrahydropapaverolone (50-100 microM). The blocking effects on membrane GCl of 4-beta-PDB and its antagonism by the inhibitors used support the concept of the involvement of PKC in regulating Cl channels of mammalian skeletal muscle fibres.

Electrogenic bicarbonate secretion in the turtle bladder: apical membrane conductance characteristics.

We have recently shown that stimulation of electrogenic HCO3- secretion is accompanied by a simultaneous increase in short-circuit current (Isc, equivalent to HCO3- secretion rate under these conditions), apical membrane capacitance (Ca, proportional to membrane area), and apical membrane conductance (Ga, proportional to membrane ionic permeability). The current experiments were undertaken to explore the ionic basis for the increase in Ga and the possibility that the rate of electrogenic HCO3- secretion is regulated by changes in Ga. Membrane electrical parameters were measured using impedance-analysis techniques before and after stimulation of electrogenic HCO3- secretion with cAMP in three solutions which contained different chloride concentrations. In another series of experiments, the effects of an anion channel blocker, anthracene-9-carboxylic acid (9-AA), were measured after stimulation of electrogenic HCO3- secretion with cAMP. The major conclusions are: (i) a measurable apical Cl- conductance exists in control hemibladders; (ii) the transport-associated increase in Ga includes a Cl(-)-conductive component; (iii) Ga also appears to reflect a HCO3- conductance; (iv) the relative magnitudes of the apical membrane conductances to Cl- and HCO3- are similar; (v) 9-AA reduces Ga and Isc in cAMP-stimulated hemibladders; and (vi) alterations in Isc appear to be mediated by changes in Ga.

Non-linear Dependence of the Dielectric Properties of Chick Embryo Myoblast Membranes Exposed to A Sinusoidal 50 Hz Magnetic Field

Primary chick embryo myoblasts can be a useful tool for studying the developmental events which accompany myoblast differentiation, particularly myoblast membrane fusion. To determine whether the electrical properties and/or fusion in these systems are affected by 50 Hz magnetic fields, chick embryo myoblast cultures were exposed to B-field intensities ranging from 1 to 10 mT. The electrical parameters of the myoblasts, i.e. membrane conductivity, membrane permittivity and the conductivity of the cell interior (cytosol) were determined by the analysis of conductivity dispersion data in the radio frequency range (10 kHz-100 MHz). Preliminary results indicate that the time of fusion (60 h) is not affected by these fields, but that the absolute values of the two membrane electrical parameters are affected. In particular, a B-field intensity-dependent decrease was observed. The maximum effect resulted after a 1 h exposure to a magnetic flux density of about 5 mT. The conductivity of the cytosol remained unchanged. These data seem to indicate that exposure to 50 Hz magnetic fields affects both static and dynamic membrane properties in primary chick embryo myoblasts.

Modulation of rat skeletal muscle chloride channels by activators and inhibitors of protein kinase C

The membrane electrical parameters and component conductances of rat extensor digitorum longus muscle fibres were studied in vitro at 30 °C with standard two microelectrode square pulse cable analysis in the presence of protein kinase C (PKC) activators and inhibitors. The PKC activator, 4-β-phorbol-12,13 dibutyrate (4-β-PDB), (2–90nM) blocked up to 67% chloride conductance (GCl) in rat skeletal muscle fibres and induced myotonic hyperexcitability. The concentration necessary to produce a 50% block of the membrane GCl was 23 nM. The “inactive” 4-α-phorbol-12,13 dibutyrate had no effect at 2 μM. The blocking effect of 4-β-PDB on GCl was prevented by preincubation of the preparations with the PKC inhibitors, staurosporine (1–5 μM) and tetrahydropapaverolone (50–100 μM). The blocking effects on membrane GCl of 4-β-PDB and its antagonism by the inhibitors used support the concept of the involvement of PKC in regulating Cl channels of mammalian skeletal muscle fibres.

Dose-dependent Effects of Ionizing Radiation onin VitroMyoblast Fusion

We have recently demonstrated by dielectric relaxation studies in the radio-frequency range that there is a sharp decrease in the conductivity and permittivity of the membranes of chick embryo myoblasts in vitro at the time of fusion (60 h) (Bonincontro et al. 1987). This sharp fall in membrane electrical parameters was subsequently shown to be due to changes in ionic flux, particularly of the Na+/K+ equilibrium (Santini et al. 1988). Ionizing radiation induces a wide variety of effects on biological membranes, including variations in membrane ionic transport. We wished to investigate if sublethal doses of gamma-irradiation could affect membrane electrical parameters and thus myoblast membrane fusion. Consequently, chick embryo myoblast aggregate cultures were irradiated with 3.25, 5.15 or 6.35 Gy at 24 h of culture. We found that the lower dose delays membrane fusion by about 10 h while the two higher doses block fusion up to 120 h of culture. Aggregates showed a very high cell viability. The possible mechanisms by which ionizing radiation causes these variations in myoblast membrane electrical properties and fusion are discussed.

Passive electrical properties of biological cell membranes determined from Maxwell—Wagner conductivity dispersion measurements

The membrane electrical parameters of erythrocyte cells in physiological saline solutions have been evaluated from conductivity measurements at radiowave frequencies, by means of appropriate mixture equations. The dielectric model adopted takes into account the main structural characteristics of the membrane structure and is able to demonstrate the contributions of the polar head groups and the hydrophobic layers. The results show the effect of some metal alkali ions (Cs +, K +, Na +, Li +) on the transport processes across the erythrocyte membrane.