The Effect of the Nonlinearity of the Response of Lipid Membranes to Voltage Perturbations on the Interpretation of Their Electrical Properties. A New Theoretical Description

Our understanding of the electrical properties of cell membranes is derived from experiments where the membrane is exposed to a perturbation (in the form of a time-dependent voltage or current change) and information is extracted from the measured output. The interpretation of such electrical recordings consists in finding an electronic equivalent that would show the same or similar response as the biological system. In general, however, there is no unique circuit configuration, which can explain a single electrical recording and the choice of an electric model for a biological system is based on complementary information (most commonly structural information) of the system investigated. Most of the electrophysiological data on cell membranes address the functional role of protein channels while assuming that the lipid matrix is an insulator with constant capacitance. However, close to their melting transition the lipid bilayers are no inert insulators. Their conductivity and their capacitance are nonlinear functions of both voltage, area and volume density. This has to be considered when interpreting electrical data. Here we show how electric data commonly interpreted as gating currents of proteins and inductance can be explained by the nonlinear dynamics of the lipid matrix itself

Solitary electromechanical pulses in Lobster neurons

Investigations of nerve activity have focused predominantly on electrical phenomena. Nerves, however, are thermodynamic systems, and changes in temperature and in the dimensions of the nerve can also be observed during the action potential. Measurements of heat changes during the action potential suggest that the nerve pulse shares many characteristics with an adiabatic pulse. First experiments in the 1980s suggested small changes in nerve thickness and length during the action potential. Such findings have led to the suggestion that the action potential may be related to electromechanical solitons traveling without dissipation. However, they have been no modern attempts to study mechanical phenomena in nerves. Here, we present ultrasensitive AFM recordings of mechanical changes on the order of 2 - 12 {\AA} in the giant axons of the lobster. We show that the nerve thickness changes in phase with voltage change. When stimulated at opposite ends of the same axon, colliding action potentials pass through one another and do not annihilate. These observations are consistent with a mechanical interpretation of the nervous impulse.Comment: 9 pages, 4 figure

Reply to "Comment on 'Penetration of Action Potentials During Collision in the Median and Lateral Giant Axons of Invertebrates'"

Berg et al. did not reproduce our results but worked on different preparations and, in one central experiment, used a significantly different electrode configuration. To clarify the situation, we have repeated their experiment on the walking leg of a lobster using an apparatus that can produce both electrode configurations. With the configuration used by Berg et al., the signal of the nerve pulse disappears when forced to pass through the region strongly perturbed by the second stimulus. In our original collision setup, pulses do not travel through perturbed regions, and pulses pass through each other without annihilation as previously reported. These results demonstrate that we handle the preparations correctly. Furthermore, they call for a reinterpretation of the so-called collision block experiment performed by Berg et al. Most likely, their results merely indicate inhibition of the nerve pulse by a strong stimulus and not annihilation upon collision as claimed

Non-invasive detection of animal nerve impulses with an atomic magnetometer operating near quantum limited sensitivity

Magnetic fields generated by human and animal organs, such as the heart, brain and nervous system carry information useful for biological and medical purposes. These magnetic fields are most commonly detected using cryogenically-cooled superconducting magnetometers. Here we present the frst detection of action potentials from an animal nerve using an optical atomic magnetometer. Using an optimal design we are able to achieve the sensitivity dominated by the quantum shot noise of light and quantum projection noise of atomic spins. Such sensitivity allows us to measure the nerve impulse with a miniature room-temperature sensor which is a critical advantage for biomedical applications. Positioning the sensor at a distance of a few millimeters from the nerve, corresponding to the distance between the skin and nerves in biological studies, we detect the magnetic field generated by an action potential of a frog sciatic nerve. From the magnetic field measurements we determine the activity of the nerve and the temporal shape of the nerve impulse. This work opens new ways towards implementing optical magnetometers as practical devices for medical diagnostics.Comment: Main text with figures, and methods and supplementary informatio

Tethered Lipid Membranes as a Nanoscale Arrangement towards Non-Invasive Analysis of Acute Pancreatitis

Extracellular heat shock proteins (HSPs) mediate immunological functions and are involved in pathologies such as infection, stress, and cancer. Here, we demonstrated the dependence of an amount of HSP70 and HSP90 in serum vs. severity of acute pancreatitis (AP) on a cohort of 49 patients. Tethered bilayer lipid membranes (tBLMs) have been developed to investigate HSPs’ interactions with tBLMs that can be probed by electrochemical impedance spectroscopy (EIS). The results revealed that HSP70 and HSP90 interact via different mechanisms. HSP70 shows the damage of the membrane, while HSP90 increases the insulation properties of tBLM. These findings provide evidence that EIS offers a novel approach for the study of the changes in membrane integrity induced by HSPs proteins. Herein, we present an alternative electrochemical technique, without any immunoprobes, that allows for the monitoring of HSPs on nanoscaled tBLM arrangement in biologics samples such us human urine. This study demonstrates the great potential of tBLM to be used as a membrane based biosensor for novel, simple, and non-invasive label-free analytical system for the prediction of AP severity

The Impact of an Anchoring Layer on the Formation of Tethered Bilayer Lipid Membranes on Silver Substrates

Tethered bilayer lipid membranes (tBLMs) have been known as stable and versatile experimental platforms for protein–membrane interaction studies. In this work, the assembly of functional tBLMs on silver substrates and the effect of the molecular chain-length of backfiller molecules on their properties were investigated. The following backfillers 3-mercapto-1-propanol (3M1P), 4-mercapto-1-butanol (4M1B), 6-mercapto-1-hexanol (6M1H), and 9-mercapto-1-nonanol (9M1N) mixed with the molecular anchor WC14 (20-tetradecyloxy-3,6,9,12,15,18,22 heptaoxahexatricontane-1-thiol) were used to form self-assembled monolayers (SAMs) on silver, which influenced a fusion of multilamellar vesicles and the formation of tBLMs. Spectroscopic analysis by SERS and RAIRS has shown that by using different-length backfiller molecules, it is possible to control WC14 anchor molecules orientation on the surface. An introduction of increasingly longer surface backfillers in the mixed SAM may be related to the increasing SAMs molecular order and more vertical orientation of WC14 at both the hydrophilic ethylenoxide segment and the hydrophobic lipid bilayer anchoring alkane chains. Since no clustering of WC14 alkane chains, which is deleterious for tBLM integrity, was observed on dry samples, the suitability of mixed-component SAMs for subsequent tBLM formation was further interrogated by electrochemical impedance spectroscopy (EIS). EIS showed the arrangement of well-insulating tBLMs if 3M1P was used as a backfiller. An increase in the length of the backfiller led to increased defectiveness of tBLMs. Despite variable defectiveness, all tBLMs responded to the pore-forming cholesterol-dependent cytolysin, vaginolysin in a manner consistent with the functional reconstitution of the toxin into phospholipid bilayer. This experiment demonstrates the biological relevance of tBLMs assembled on silver surfaces and indicates their utility as biosensing elements for the detection of pore-forming toxins in liquid samples