Changes In Transmembrane Voltage Research Articles

Genetically engineered HEK cells as a valuable tool for studying electroporation in excitable cells

Electric pulses used in electroporation-based treatments have been shown to affect the excitability of muscle and neuronal cells. However, understanding the interplay between electroporation and electrophysiological response of excitable cells is complex, since both ion channel gating and electroporation depend on dynamic changes in the transmembrane voltage (TMV). In this study, a genetically engineered human embryonic kidney cells expressing NaV1.5 and Kir2.1, a minimal complementary channels required for excitability (named S-HEK), was characterized as a simple cell model used for studying the effects of electroporation in excitable cells. S-HEK cells and their non-excitable counterparts (NS-HEK) were exposed to 100 µs pulses of increasing electric field strength. Changes in TMV, plasma membrane permeability, and intracellular Ca2+ were monitored with fluorescence microscopy. We found that a very mild electroporation, undetectable with the classical propidium assay but associated with a transient increase in intracellular Ca2+, can already have a profound effect on excitability close to the electrostimulation threshold, as corroborated by multiscale computational modelling. These results are of great relevance for understanding the effects of pulse delivery on cell excitability observed in context of the rapidly developing cardiac pulsed field ablation as well as other electroporation-based treatments in excitable tissues.

PIP2 binding at the voltage sensor domain facilitates KCNQ1 VSD activation and gating.

The cardiac KCNQ1 channel is involved in cell hyperpolarisation, and loss-of-function mutations are associated with Long QT syndrome (LQTS), which can cause sudden cardiac arrest. The KCNQ1 channel is gated by the changes in transmembrane voltage and crucially requires the binding of two phosphatidylinositol-4,5-bisphosphate (PIP2) molecules to open the channel. The recent structure of KCNQ1 has revealed a PIP2 binding site at the voltage-sensor domain (VSD). In addition, previous modelling and electrophysiological studies have proposed another PIP2 binding site at the pore domain. Using enhanced sampling molecular dynamics simulation, we show that PIP2 binding at the VSD facilitates its activation, whereas PIP2 interaction of the pore facilitates VSD-pore coupling by straightening the S6 helix. Our simulation proposes that PIP2 binding at the VSD acts as a counter charge pausing point during the voltage-dependent activation process, lowering the energy barrier during the S4 translocation. Together, our work highlights the importance of PIP2 at the VSD as an activator to the VSD activation, in addition to our previous knowledge as the key coupler between the VSD and the pore.

A red fluorescent protein with improved monomericity enables ratiometric voltage imaging with ASAP3

A ratiometric genetically encoded voltage indicator (GEVI) would be desirable for tracking transmembrane voltage changes in the presence of sample motion. We performed combinatorial multi-site mutagenesis on a cyan-excitable red fluorescent protein to create the bright and monomeric mCyRFP3, which proved to be uniquely non-perturbing when fused to the GEVI ASAP3. The green/red ratio from ASAP3-mCyRFP3 (ASAP3-R3) reported voltage while correcting for motion artifacts, allowing the visualization of membrane voltage changes in contracting cardiomyocytes and throughout the cell cycle of motile cells.

Assays of CFTR Function In Vitro, Ex Vivo and In Vivo.

Cystic fibrosis, a multi-organ genetic disease, is characterized by abnormal function of the cystic fibrosis transmembrane conductance regulator (CFTR) protein, a chloride channel at the apical membrane of several epithelia. In recent years, therapeutic strategies have been developed to correct the CFTR defect. To evaluate CFTR function at baseline for diagnosis, or the efficacy of CFTR-restoring therapy, reliable tests are needed to measure CFTR function, in vitro, ex vivo and in vivo. In vitro techniques either directly or indirectly measure ion fluxes; direct measurement of ion fluxes and quenching of fluorescence in cell-based assays, change in transmembrane voltage or current in patch clamp or Ussing chamber, swelling of CFTR-containing organoids by secondary water influx upon CFTR activation. Several cell or tissue types can be used. Ex vivo and in vivo assays similarly evaluate current (intestinal current measurement) and membrane potential differences (nasal potential difference), on tissues from individual patients. In the sweat test, the most frequently used in vivo evaluation of CFTR function, chloride concentration or stimulated sweat rate can be directly measured. Here, we will describe the currently available bio-assays for quantitative evaluation of CFTR function, their indications, advantages and disadvantages, and correlation with clinical outcome measures.

Network analysis reveals how lipids and other cofactors influence membrane protein allostery.

Many membrane proteins are modulated by external stimuli, such as small molecule binding or change in pH, transmembrane voltage, or temperature. This modulation typically occurs at sites that are structurally distant from the functional site. Revealing the communication, known as allostery, between these two sites is key to understanding the mechanistic details of these proteins. Residue interaction networks of isolated proteins are commonly used to this end. Membrane proteins, however, are embedded in a lipid bilayer, which may contribute to allosteric communication. The fast diffusion of lipids hinders direct use of standard residue interaction networks. Here, we present an extension that includes cofactors such as lipids and small molecules in the network. The novel framework is applied to three membrane proteins: a voltage-gated ion channel (KCNQ1), a G-protein coupled receptor (GPCR-β2 adrenergic receptor), and a pH-gated ion channel (KcsA). Through systematic analysis of the obtained networks and their components, we demonstrate the importance of lipids for membrane protein allostery. Finally, we reveal how small molecules may stabilize different protein states by allosterically coupling and decoupling the protein from the membrane.

Mechanisms of Light-Induced Deformations in Photoreceptors

Biological cells deform on a nanometer scale when their transmembrane voltage changes, an effect that has been visualized during the action potential using quantitative phase imaging. Similar changes in the optical path length have been observed in photoreceptor outer segments after a flash stimulus via phase-resolved optical coherence tomography. These optoretinograms reveal a fast, millisecond-scale contraction of the outer segments by tens of nanometers, followed by a slow (hundreds of milliseconds) elongation reaching hundreds of nanometers. Ultrafast measurements of the contractile response using line-field phase-resolved optical coherence tomography show a logarithmic increase in amplitude and a decreasing time to peak with increasing stimulus intensity. We present a model that relates the early receptor potential to these deformations based on the voltage-dependent membrane tension—the mechanism observed earlier in neurons and other electrogenic cells. The early receptor potential is caused by conformational changes in opsins after photoisomerization, resulting in the fractional shift of the charge across the disk membrane. Lateral repulsion of the ions on both sides of the membrane affects its surface tension and leads to its lateral expansion. Because the volume of the disks does not change on a millisecond timescale, their lateral expansion leads to an axial contraction of the outer segment. With increasing stimulus intensity and the resulting tension, the area expansion coefficient of the disk membrane also increases as thermally induced fluctuations are pulled flat, resisting further expansion. This leads to the logarithmic saturation observed in measurements as well as the peak shift in time. This imaging technique therefore relates the structural changes in the photoreceptor to the underlying neurological function of transducing light into electrical signals. Such label-free optical monitoring of neural activity using fast interferometry may be applicable not only to optoretinography but also to neuroscience in general.

Actin networks regulate the cell membrane permeability during electroporation

Transient physical disruption of cell membranes by electric pulses (or electroporation) has significance in biomedical and biological applications requiring the delivery of exogenous (bio)molecules to living cells. We demonstrate that actin networks regulate the cell membrane permeability during electroporation. Disruption of actin networks increases the uptake of membrane-impermeable molecules such as propidium iodide during electroporation. Our experiments at different temperatures ranging from 11 °C to 37 °C show that molecular uptake during electroporation increases with temperature. Furthermore, by examining the temperature-dependent kinetics of propidium iodide uptake, we infer that the activation energy barrier of electroporation is lowered when the actin networks are disrupted. Our numerical calculations of transmembrane voltage show that the reduced activation energy barrier for the cells with disrupted actin is not a consequence of the changes in transmembrane voltage associated with changes in the cell shape due to the disruption of actin, indicating that this could be due to changes in membrane mechanical properties. Our results suggest that the current theoretical models of electroporation should be advanced further by including the contributions of the cytoskeletal networks on the cell membrane permeability during the delivery of exogenous materials.

Pulsed Electric Fields Can Create Pores in the Voltage Sensors of Voltage-Gated Ion Channels

Pulsed electric fields are increasingly used in medicine to transiently increase the cell membrane permeability via electroporation to deliver therapeutic molecules into the cell. One type of event that contributes to this increase in membrane permeability is the formation of pores in the membrane lipid bilayer. However, electrophysiological measurements suggest that membrane proteins are affected as well, particularly voltage-gated ion channels (VGICs). The molecular mechanisms by which the electric field could affects these molecules remain unidentified. In this study, we used molecular dynamics simulations to unravel the molecular events that take place in different VGICs when exposing them to electric fields mimicking electroporation conditions. We show that electric fields can induce pores in the voltage-sensor domains (VSDs) of different VGICs and that these pores form more easily in some channels than in others. We demonstrate that poration is more likely in VSDs that are more hydrated and are electrostatically more favorable for the entry of ions. We further show that pores in VSDs can expand into so-called complex pores, which become stabilized by lipid headgroups. Our results suggest that such complex pores are considerably more stable than conventional lipid pores, and their formation can lead to severe unfolding of VSDs from the channel. We anticipate that such VSDs become dysfunctional and unable to respond to changes in transmembrane voltage, which is in agreement with previous electrophysiological measurements showing a decrease in the voltage-dependent transmembrane ionic currents after pulse treatment. Finally, we discuss the possibility of activation of VGICs by submicrosecond-duration pulses. Overall, our study reveals a new, to our knowledge, mechanism of electroporation through membranes containing VGICs.

The HCN channel voltage sensor undergoes a large downward motion during hyperpolarization.

Voltage-gated ion channels (VGICs) contain positively-charged residues within the S4 helix of the voltage-sensing domain (VSD) that are displaced in response to changes in transmembrane voltage, promoting conformational changes that open the pore. Pacemaker HCN channels are unique among VGICs because their open probability is increased by membrane hyperpolarization rather than depolarization. Here we measured the precise movement of the S4 helix of a sea urchin HCN channel using transition metal ion fluorescence resonance energy transfer (tmFRET). We show that the S4 undergoes a significant (~10 Å) downward movement in response to membrane hyperpolarization. Furthermore, by applying distance constraints determined from tmFRET experiments to Rosetta modeling, we reveal that the C-terminal part of the S4 helix exhibits an unexpected tilting motion during hyperpolarization activation. These data provide a long-sought glimpse of the hyperpolarized state of a functioning VSD and also a framework for understanding the dynamics of reverse gating in HCN channels.

Hybrid analytical-numerical approach for investigation of differential effects in normal and cancer cells under electroporation.

Electroporation has offered important biomedical applications in electrochemotherapy, tissue ablation and gene editing recently. Time and computation efficient analytical and numerical models should be developed to understand the differential effects of electroporation on normal and cancer cells. In this work, we present a hybrid analytical–numerical approach to investigate the behavior of normal and cancer cells under electroporation. We have compared the human breast cancer cell (MCF-7) and non-tumorigenic human breast cell (MCF-10A) under electroporation in terms of change in transmembrane voltage and pore formation on cell surface. The effects of electric pulse time, amplitude and membrane conductivity variation are analyzed in a systematic manner. To accelerate the calculation of transmembrane voltage, we have introduced a simple Multilayer Electric Potential Model (MEPM) which calculates the potential distribution across the cell analytically. The MEPM calculates electric potential distribution across a biological cell sandwiched between two semi-circular electrodes held at fixed potential, by solving the Laplace's equation over an equivalent planar, multilayer geometry. The MEPM model is then used in a Finite Element Method (FEM) based numerical model of electroporation. Transmembrane voltage and pore density for electroporated MCF-10A are estimated to be 1.31 V and 2.98 × 1013 m−2 respectively, and for MCF-7 the estimated values are 0.53 V and 1.93 × 1014 m−2, respectively. Our results suggest that under electroporation, the cancer cell's membrane get much more permeabilized than its counterpart normal cell even at small values of transmembrane voltage. This work provides a theoretical basis for further experimental exploration of electroporation process in cancer therapy, and serves as a design tool for performance optimization.

Towards systematic, data-driven validation of a collaborative, multi-scale model of Caenorhabditis elegans.

The OpenWorm Project is an international open-source collaboration to create a multi-scale model of the organism Caenorhabditis elegans At each scale, including subcellular, cellular, network and behaviour, this project employs one or more computational models that aim to recapitulate the corresponding biological system at that scale. This requires that the simulated behaviour of each model be compared with experimental data both as the model is continuously refined and as new experimental data become available. Here we report the use of SciUnit, a software framework for model validation, to attempt to achieve these goals. During project development, each model is continuously subjected to data-driven 'unit tests' that quantitatively summarize model-data agreement, identifying modelling progress and highlighting particular aspects of each model that fail to adequately reproduce known features of the biological organism and its components. This workflow is publicly visible via both GitHub and a web application and accepts community contributions to ensure that modelling goals are transparent and well-informed.This article is part of a discussion meeting issue 'Connectome to behaviour: modelling C. elegans at cellular resolution'.

No voltage change at skeletal muscle SR membrane during Ca2+ release-just Mermaids on acid.

Calcium ions control multiple physiological functions by binding to extracellular and intracellular targets. One of the best-studied Ca2+-dependent functions is contraction of smooth and striated muscle tissue, which results from Ca2+ ligation to calmodulin and troponin C, respectively. Ca2+ signaling typically involves flux of the ion across membranes via specifically gated channel proteins. Because calcium ions are charged, they possess the ability to generate changes in the respective transmembrane voltage. Ca2+-dependent voltage alterations of the surface membrane are easily measured using microelectrodes. A well-known example is the characteristic plateau phase of the action potential in cardiac ventricular cells that results from the opening of voltage-gated L-type Ca2+ channels. Ca2+ ions are also released from intracellular storage compartments in many cells, but these membranes are not accessible to direct voltage recording with microelectrodes. In muscle, for example, release of Ca2+ from the sarcoplasmic reticulum (SR) to the myoplasm constitutes a flux that is considerably larger than the entry flux from the extracellular space. Whether this flux is accompanied by a voltage change across the SR membrane is an obvious question of mechanistic importance and has been the subject of many investigations. Because the tiny spaces enclosed by the SR membrane are inaccessible to microelectrodes, alternative methods have to be applied. In a study by Sanchez et al. (2018. J. Gen. Physiol. https://doi.org/10.1085/jgp.201812035) in this issue, modern confocal light microscopy and genetically encoded voltage probes targeted to the SR were applied in a new approach to search for changes in the membrane potential of the SR during Ca2+ release.

An Introduction to Computational Modeling of Cardiac Electrophysiology and Arrhythmogenicity.

Mathematical modeling is a powerful tool to study the complex and orchestrated biological process of cardiac electrical activity. By integrating experimental data from key components of cardiac electrophysiology, systems biology simulations can complement empirical findings, provide quantitative insight into physiological and pathophysiological mechanisms of action, and guide new hypotheses to better understand this complex biological system to develop novel cardiotherapeutic approaches. In this chapter, we briefly introduce in silico methods to describe the dynamics of physiological and pathophysiological single-cell and tissue-level cardiac electrophysiology. Using a "bottom-up" approach, we first describe the basis of ion channel mathematical models. Next, we discuss how the net flux of ions through such channels leads to changes in transmembrane voltage during cardiomyocyte action potentials. By applying these fundamentals, we describe how action potentials propagate in models of cardiac tissue. In addition, we provide case studies simulating single-cell and tissue-level arrhythmogenesis, as well as promising approaches to circumvent or overcome such adverse events. Overall, basic concepts and tools are discussed in this chapter as an accessible introduction to nonmathematicians to foster an understanding of electrophysiological modeling studies andhelpfacilitate communication with dry lab colleagues and collaborators.

Electrical coupling between ventricular myocytes and myofibroblasts in the infarcted mouse heart.

Recent studies have demonstrated electrotonic coupling between scar tissue and the surrounding myocardium in cryoinjured hearts. However, the electrical dynamics occurring at the myocyte-nonmyocyte interface in the fibrotic heart remain undefined. Here, we sought to develop an assay to interrogate the nonmyocyte cell type contributing to heterocellular coupling and to characterize, on a cellular scale, its voltage response in the infarct border zone of living hearts. We used two-photon laser scanning microscopy in conjunction with a voltage-sensitive dye to record transmembrane voltage changes simultaneously from cardiomyocytes and adjoined nonmyocytes in Langendorff-perfused mouse hearts with healing myocardial infarction. Transgenic mice with cardiomyocyte-restricted expression of a green fluorescent reporter protein underwent permanent coronary artery ligation and their hearts were subjected to voltage imaging 7-10 days later. Reporter-negative cells, i.e. nonmyocytes, in the infarct border zone exhibited depolarizing transients at a 1:1 coupling ratio with action potentials recorded simultaneously from adjacent, reporter-positive ventricular myocytes. The electrotonic responses in the nonmyocytes exhibited slower rates of de- and repolarization compared to the action potential waveform of juxtaposed myocytes. Voltage imaging in infarcted hearts expressing a fluorescent reporter specifically in myofibroblasts revealed that the latter were electrically coupled to border zone myocytes. Their voltage transient properties were indistinguishable from those of nonmyocytes in hearts with cardiomyocyte-restricted reporter expression. The density of connexin43 expression at myofibroblast-cardiomyocyte junctions was ∼5% of that in the intercalated disc regions of paired ventricular myocytes in the remote, uninjured myocardium, whereas the ratio of connexin45 to connexin43 expression levels at heterocellular contacts was ∼1%. Myofibroblasts contribute to the population of electrically coupled nonmyocytes in the infarct border zone. The slower kinetics of myofibroblast voltage responses may reflect low electrical conductivity across heterocellular junctions, in accordance with the paucity of connexin expression at myofibroblast-cardiomyocyte contacts.

Exploring Instructive Physiological Signaling with the Bioelectric Tissue Simulation Engine.

Bioelectric cell properties have been revealed as powerful targets for modulating stem cell function, regenerative response, developmental patterning, and tumor reprograming. Spatio-temporal distributions of endogenous resting potential, ion flows, and electric fields are influenced not only by the genome and external signals but also by their own intrinsic dynamics. Ion channels and electrical synapses (gap junctions) both determine, and are themselves gated by, cellular resting potential. Thus, the origin and progression of bioelectric patterns in multicellular tissues is complex, which hampers the rational control of voltage distributions for biomedical interventions. To improve understanding of these dynamics and facilitate the development of bioelectric pattern control strategies, we developed the BioElectric Tissue Simulation Engine (BETSE), a finite volume method multiphysics simulator, which predicts bioelectric patterns and their spatio-temporal dynamics by modeling ion channel and gap junction activity and tracking changes to the fundamental property of ion concentration. We validate performance of the simulator by matching experimentally obtained data on membrane permeability, ion concentration and resting potential to simulated values, and by demonstrating the expected outcomes for a range of well-known cases, such as predicting the correct transmembrane voltage changes for perturbation of single cell membrane states and environmental ion concentrations, in addition to the development of realistic transepithelial potentials and bioelectric wounding signals. In silico experiments reveal factors influencing transmembrane potential are significantly different in gap junction-networked cell clusters with tight junctions, and identify non-linear feedback mechanisms capable of generating strong, emergent, cluster-wide resting potential gradients. The BETSE platform will enable a deep understanding of local and long-range bioelectrical dynamics in tissues, and assist the development of specific interventions to achieve greater control of pattern during morphogenesis and remodeling.

Membrane resistance and shunting inhibition: where biophysics meets state-dependent human neurophysiology.

Activation of neurons not only changes their membrane potential and firing rate but as a secondary action reduces membrane resistance. This loss of resistance, or increase of conductance, may be of central importance in non‐invasive magnetic or electric stimulation of the human brain since electrical fields cause larger changes in transmembrane voltage in resting neurons with low membrane conductances than in active neurons with high conductance. This may explain why both the immediate effects and after‐effects of brain stimulation are smaller or even reversed during voluntary activity compared with rest. Membrane conductance is also increased during shunting inhibition, which accompanies the classic GABAA IPSP. This short‐circuits nearby EPSPs and is suggested here to contribute to the magnitude and time course of short‐interval intracortical inhibition and intracortical facilitation.

Internal Sodium in GPCRs Strongly Responds to Transmembrane Voltage Changes

G protein-coupled receptors (GPCRs) are the largest superfamily of membrane proteins in the human genome, mediating the propagation of extracellular ligand binding information into intracellular signal transduction cascades. Crystal structures have revealed a water-filled hydrophilic internal pocket within their transmembrane domain, extending from the orthosteric ligand-binding site to regions near the G protein binding site. Recent high-resolution structures have identified a sodium ion near the base of this pocket, coordinated by highly conserved residues (1,2). Owing to this conservation level, sodium binding may play a role for most rhodopsin-like GPCRs (3). Recently, functional or conformational effects have been shown to be elicited by physiological transmembrane voltage in several GPCRs, and voltage-clamp recordings have determined a gating charge of ∼0.85 e in the M1 and M2 muscarinic receptors (4). However, the nature of the voltage sensor has remained enigmatic. Here we show by MD and computational electrophysiology simulations (5) that the internal sodium ion in the delta-opioid and muscarinic receptors is highly mobile under the influence of transmembrane voltage changes. By calculating the energetics of ion movement along the pocket axis, we find that the free energy barriers to migration can be overcome by the physiological voltage range. Furthermore, we demonstrate that the motion along the axis creates a gating charge in excellent agreement with the experimentally measured gating currents. Thus, voltage-induced movements of Na+ in GPCRs may represent an attractive mechanism for voltage-regulation of GPCRs.1. Liu, W. et al., Science 337: 232-236, 2012.2. Fenalti, G. et al., Nature 506: 191-196, 2014.3.Katritch, V. et al., TiBS 39: 233-244, 2014.4. Ben-Chaim, Y. et al., Nature 444: 106-9, 2006.5. Kutzner, C. et al., Biophysical Journal 101: 809-817, 2011.

Selective Cation Binding to the Gating-Ring Triggers Independent RCK Motions in the BK Channel

Large conductance voltage and calcium dependent potassium channels (BK channels) are important controllers of cell excitability. Functional BK channels are formed as tetramers of alpha subunits. Their open probability is regulated synergistically by changes in transmembrane voltage and intracellular concentration of divalent cations including Ca2+, Mg2+, Cd2+ and Ba2+. The voltage sensor resides within the transmembrane region of the alpha subunit, whereas a large C-terminal intracellular region called ‘gating ring’ senses divalent cation binding. The gating ring assembles as a large tetrameric structure that contains a total of eight Regulator of Conductance of K (RCK) domains, two from each subunit. In each RCK domain resides one high-affinity Ca2+ binding site. Even though the two RCK domains of each subunit (RCK1 and RCK2) are similar, their Ca2+ binding sites are not, particularly in their selectivity to divalent ions other than Ca2+. For example, Cd2+ effect is mediated exclusively through the RCK1 domains, while Ba2+ and Mg2+ (at low concentrations) prefer solely the site from the RCK2 domains. Using patch clamp fluorometry, we have shown previously that large rearrangements of the gating-ring occur in response to Ca2+ and voltage. By themselves, Cd2+, Mg2+ and Ba2+ induce movements of the gating-ring that are substantially smaller than those triggered by Ca2+. Full extent motions are achieved once binding sites from RCK1 and RCK2 domains are occupied by Cd2+ and Ba2+, or Cd2+ and Mg2+. These results suggest that the gating ring structure is flexible, potentially allowing independent movements of the RCK1 and RCK2 domains.

Voltage Dependence of BK Channels Gating Ring Motion Studied by State Dependent FRET

Large conductance voltage- and calcium-dependent potassium channels (BK) open probability is regulated by changes in transmembrane voltage and intracellular divalent ions (Ca2+ and Mg2+) concentration. The gating ring, formed by the two RCK (Regulator of Conductance of Potassium) domains present in the C-terminal of each subunit of the tetramer, contains the divalent ion binding sites. We previously have showed that large rearrangements of the gating ring occur in a Ca2+ and voltage dependent manner. This movement of the gating ring is not directly related with the opening of the channel. It appears that a change in the voltage dependence of the open probability of the channel does not modify the voltage dependence of the movement of the gating ring. However, the voltage dependence of the movement of the gating ring is shifted toward a more negative potential if we modify the voltage dependence of the voltage sensor by co-expression of the BK alpha subunit with the Beta1 subunit or by mutations. These results indicate that the voltage dependent movement of the gating ring appears to be related to the voltage dependence of the voltage sensor and not directly related with the opening of the gate.

Electromechanical properties of biomembranes and nerves

Lipid membranes are insulators and capacitors, which can be charged by an external electric field. This phenomenon plays an important role in the field of electrophysiology, for instance when describing nerve pulse conduction. Membranes are also made of polar molecules meaning that they contain molecules with permanent electrical dipole moments. Therefore, the properties of membranes are subject to changes in trans-membrane voltage. Vice versa, mechanical forces on membranes lead to changes in the membrane potential. Associated effects are flexoelectricity, piezoelectricity, and electrostriction.Lipid membranes can melt from an ordered to a disordered state. Due to the change of membrane dimensions associated with lipid membrane melting, electrical properties are linked to the melting transition. Melting of the membrane can induce changes in trans-membrane potential, and application of voltage can lead to a shift of the melting transition. Further, close to transitions membranes are very susceptible to piezoelectric phenomena.We discuss these phenomena in relation with the occurrence of lipid ion channels. Close to melting transitions, lipid membranes display step-wise ion conduction events, which are indistinguishable from protein ion channels. These channels display a voltage-dependent open probability. One finds asymmetric current-voltage relations of the pure membrane very similar to those found for various protein channels. This asymmetry falsely has been considered a criterion to distinguish lipid channels from protein channels. However, we show that the asymmetry can arise from the electromechanical properties of the lipid membrane itself.Finally, we discuss electromechanical behavior in connection with the electromechanical theory of nerve pulse transduction. It has been found experimentally that nerve pulses are related to changes in nerve thickness. Thus, during the nerve pulse a solitary mechanical pulse travels along the nerve. Due to electromechanical coupling it is unavoidable that this pulse generates a trans-membrane voltage. In the past, we have proposed that this electromechanical pulse is the origin of the action potential in nerves.