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Electroreception in G carapo: detection of changes in waveform of the electrosensory signals.

Electric fish evaluate the near environment by detecting changes in their self-generated electric organ discharge. To investigate impedance modulation of the self-generated electric field, this field was measured at the electrosensory fovea of Gymnotus carapo in the presence and absence of objects. Changes in local fields provoked by resistive objects were predicted by the change in total energy. Objects with capacitive impedance generated large variations in the relative importance of the different waveform components of the electric organ discharge. We tested the hypothesis that fish discriminate changes in waveform as well as increases in total energy using the novelty response, which is a behavioural response consisting of a transient acceleration of EOD frequency that can follow a change in object impedance. For resistive loads, the amplitude of novelty responses was well predicted by the increase in total energy. For complex loads, the amplitude of novelty responses was correlated not only with increases in total energy but also with waveform changes, consisting of reductions in the early slow negative wave and increases in the late sharp negative wave. The total energy and waveform effects appeared to be additive. These results indicate that G. carapo discriminates complex impedance based on an evaluation of different waveform parameters.

Animals↗

Impedance analysis and single-channel recordings on nano-black lipid membranes based on porous alumina.

Ordered porous alumina substrates with pore diameters of 55 and 280 nm, respectively, were produced and utilized as a support to prepare membranes suspending the pores of the material. Highly ordered porous alumina was prepared by an anodization process followed by dissolution of the remaining aluminum and alumina at the backside of the pores. The dissolution process of Al(2)O(3) at the backside of the pores was monitored by electrical impedance spectroscopy ensuring the desired sieve-like structure of the porous alumina. One side of the porous material with an area of 7 mm(2) was coated with a thin gold layer followed by chemisorption of 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol. The hydrophobic monolayer on top of the upper surface was a prerequisite for the formation of suspending membranes, termed nano-black lipid membranes (nano-BLMs). The formation process, and long-term and mechanical stability of the nano-BLMs were followed by electrical impedance spectroscopy indicating the formation of lipid bilayers with typical specific membrane capacitances of (0.65 +/- 0.2) micro F/cm(2) and membrane resistances of up to 1.6 x 10(8) Omega cm(2). These high membrane resistances allowed for single-channel recordings. Gramicidin as well as alamethicin was successfully inserted into the nano-BLMs exhibiting characteristic conductance states.

Adsorption↗

Computer simulation of action potentials and afterpotentials in mammalian myelinated axons: the case for a lower resistance myelin sheath.

Depolarizing afterpotentials, recorded in peripheral nerves [Barrett and Barrett (1982) J. Physiol., Lond. 323, 117-144] and spinal axons [Blight and Someya (1985) Neuroscience 15, 1-12], have been interpreted as representing passive discharge of axolemmal capacitance. This interpretation requires a lower resistance pathway through the myelin sheath than previous measurements have suggested. A computer model was used to examine the contribution of the electrical characteristics of nerve fibers to action potential conduction and afterpotential generation. The model consisted of a resistance-capacitance network representing a chain of 20 internodes. The resistances of node, internode and myelin sheath, deduced from observations in the accompanying paper, [Blight and Someya (1985) Neuroscience, 15] were found to produce suitable length and time constants, and prolonged afterpotentials, when inserted into the model. Similar length and time constants were found using a conventional model of the axon, based on measurements from isolated peripheral fibers, but this did not reproduce the afterpotentials. Action-potential conduction velocity is enhanced by reducing the time constant and increasing the length constant. The problem of minimizing the internodal time constant was met in the conventional model through the low parallel resistance of the node, while in the new model it was met by reducing the resistance of the myelin sheath. The latter strategy required the nodal leakage resistance to be higher than values from single fiber measurements (ca 250 M omega rather than ca 50 M omega) in order to maintain the length constant similar to the conventional model. Simulation of the recorded potentials required the resistance of the myelin lamellae to be approx. 100 omega cm2. The model quantitatively reproduced the voltage response of the axon to injected current pulses and to propagated action potentials, using Frankenhaeuser-Huxley kinetics. [Frankenhaeuser and Huxley (1964) J. Physiol., Lond. 171, 302-315; Frankenhaeuser and Moore (1963) J. Physiol., Lond. 169, 431-437]. The short duration components of the afterpotential, observed in mammalian recordings were reproduced by assuming a leakage pathway in the myelin sheath, at the impalement site. The calculated lower resistance of the myelin sheath was such that it minimized the effective internodal time constant for a given nodal resistance. This appears to free the myelinated fiber from the alternative requirement for a high nodal leakage conductance.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

Impedance and QCM analysis of the protein resistance of self-assembled PEGylated alkanethiol layers on gold.

In this study, we describe the formation and characterization of self-assembled layers of undecanthiol with poly(ethylene glycol) (PEG) moieties of 350Da (PEG350-undecanthiol) and 2000Da (PEG2000-undecanthiol) on gold surfaces. The functionalized surfaces were investigated by means of electrical impedance spectroscopy, which allows calculating the surface coverage from the obtained capacitance values of the formed layers. For both, PEG350-undecanthiol and PEG2000-undecanthiol layers, a surface coverage well above 90% was obtained. Protein resistance of those layers was investigated using the quartz crystal microbalance (QCM) technique, which enables one to monitor protein adsorption label free and in a time resolved manner. The change in resonance frequency of the quartz plate was monitored upon addition of fetal bovine serum indicating that PEG-functionalized surfaces are partly protein resistant compared to hydroxyundecanthiol- and non-functionalized gold surfaces. From QCM experiments, where only a single protein component was added to the PEG-functionalized gold surface, we conclude that the surfaces are fully resistant against serum albumins, while the main protein components that adsorb are globulins. A kinetic analysis reveals that PEG modified gold surfaces do not only significantly diminish the overall amount of bound protein but also significantly slows down the adsorption process.

Adsorption↗

Electrical impedance, ultrastructure and ion transport in foetal gastric mucosa.

1. The voltage transient developed across rabbit foetal gastric mucosa in response to rectangular current pulses was measured under in vitro conditions. The short-circuit current was also measured.2. The voltage transients were used to calculate equivalent circuit parameters. The mucosa could be represented as two large capacitances in series, each shunted by a high conductance.3. Electron microscopy was carried out on some preparations after the electrical measurements had been made. An extensive smooth membrane tubular system was observed on the apical sides of the cells, with openings into the gastric lumen.4. Anoxia resulted in a reversible reduction in value of the large capacitance and short-circuit current: the relative changes in these two quantities were very closely linked. Electron microscopy showed a large and reversible decline in the extent of the apical tubular system during anoxia.5. It was concluded that the apical tubular system plays an important role in ion transport in this preparation. Considerations of the magnitude of the capacitances and of membrane area suggest that the smaller capacitor represents the plasma membrane. The dieletric constant of the smooth membrane of the tubular system is an order of magnitude lower than that of the plasma membrane.

Animals↗

An electrically isolated balanced wideband current source: basic considerations and design.

At relatively high frequencies, the application of an alternating current through the body or a body segment results in electromagnetic stray fields which reduce the amount of current actually injected into the tissue under study. This radiation effect can be reduced by use of a symmetrical configuration current source. The symmetry of such an arrangement, however, depends on the stray capacitances of the source with respect to surrounding equipment. To minimise these effects, it is required that the source is electrically isolated from the surrounding equipment and the subject under study. In this manner stray capacitances with respect to elements of the current source are reduced. In such a configuration common mode voltages to the input amplifier of the measuring system are also reduced. The paper describes design considerations and the implementation of a wideband current source capable of injecting alternating current in the order of 300 microARMS into biological tissue having impedances up to 1 k omega. Current stabilisation is obtained by means of a control circuit which measures the actual current passing through the tissue under study. Leakage currents arising from shielding and stray capacitances are compensated for. The usable frequency range is between 4 kHz and 1024 kHz and current stability is better than 0.2%. Through the use of a symmetrical, floating circuit a configuration is obtained which substantially reduces stray effects. The current source is connected to other circuits by means of two isolation ports: (1) a transformer coupling for the carrier frequency; and (2) an opto-coupler to transfer a phase reference signal obtained from current measurement. The current amplitude can be modulated by controlling the reference input to the control loop by means of a third auxiliary isolation port for transfer of the modulating signal.

Cardiography, Impedance↗

[The electrical resistance and impedance of mammalian muscles during the first days after slaughter. I: bovine diaphragm].

In the view of characterizing the animal tissues with respect to their physical properties, the AA. have investigated on the behaviour of the electrical responses of bovine diaphragmatic muscle. In particolar the resistance and the impedance (at a fixed frequency of 20 Hz, sinusoidal wave) have been checked in samples from the 4th up to the 148th hour after slaughter. The results of these experiments show that, during the maturation of the meat, there is a decrease of the values of the resistance and of the capacitive reactance. On the basis of the data acquired, the only hypothesis explaining the course of the research could be that the electrical properties of the meat are greatly influenced in a very relevant way by capacitive structures and in lower degree by a pure resistive medium.

Animals↗

A method and medium for the electrical detection of Listeria spp. from food.

The development of a liquid medium for the detection of Listeria spp. by capacitance monitoring of food samples previously enriched in UVM 1 broth is described. Rapid growth of Listeria monocytogenes was shown to occur in liquid media with selectivity based on antibiotics found in Oxford agar. The final capacitance medium contained higher concentrations of the Oxford selective agents than Oxford agar and did not require the esculin/ferric ammonium citrate reaction to be observed. The medium relied upon the ability of Listeria spp. to induce a greater than 30% change in capacitance within 30 h. When run in parallel with the Listeria spp. test samples of a large food company, the method gave far fewer false-positive results than Fraser broth.

Bacteriological Techniques↗

Chronic lead exposure reduces junctional resistance at an electrical synapse.

Both acute and chronic lead exposure have been found to inhibit transmission at chemical synapses, possibly by interfering with inward calcium current. We have found that chronic lead exposure slightly reduces input resistance and greatly reduces the junctional resistance between two strongly electrically coupled neurons in the pond snail Lymnaea stagnalis. The net effect is to increase the strength of electrical coupling. A reduction in gap junctional resistance would also be expected to increase the flow of small molecules between cells. However, Lucifer Yellow injections did not reveal dye-coupling between the cells. Lead exposure also increases the capacitance of the neurons.

Animals↗

Direct measurement of specific membrane capacitance in neurons.

The specific membrane capacitance (C(m)) of a neuron influences synaptic efficacy and determines the speed with which electrical signals propagate along dendrites and unmyelinated axons. The value of this important parameter remains controversial. In this study, C(m) was estimated for the somatic membrane of cortical pyramidal neurons, spinal cord neurons, and hippocampal neurons. A nucleated patch was pulled and a voltage-clamp step was applied. The exponential decay of the capacitative charging current was analyzed to give the total membrane capacitance, which was then divided by the observed surface area of the patch. C(m) was 0.9 microF/cm(2) for each class of neuron. To test the possibility that membrane proteins may alter C(m), embryonic kidney cells (HEK-293) were studied before and after transfection with a plasmid coding for glycine receptor/channels. The value of C(m) was indistinguishable in untransfected cells and in transfected cells expressing a high level of glycine channels, indicating that differences in transmembrane protein content do not significantly affect C(m). Thus, to a first approximation, C(m) may be treated as a "biological constant" across many classes of neuron.

Animals↗

Membrane changes associated with the temperature-sensitive P85gag-mos-dependent transformation of rat kidney cells as determined by dielectrophoresis and electrorotation.

Conventional dielectrophoresis (cDEP) and electrorotation (ROT) measurements have been used to determine the dielectric properties of a clone of normal rat kidney cells, designated 6m2, that exhibits a transformed phenotype at 33 degrees C and a non-transformed phenotype at 39 degrees C. cDEP measurements of the crossover frequencies at which individual 6m2 cells experienced zero cDEP force performed as a function of the conductivity of the suspension medium revealed that, in response to a temperature shift from 33 degrees C to 39 degrees C for 24 h, the mean specific cell membrane capacitance and conductance fell significantly (P < 0.01) from 42.3 (+/-1.3) to 30.3 (+/-2.9) mF/m2 and 743 (+/-422) to 567 (+/-326) S/m2, respectively. ROT analyses demonstrated a similar reduction for the membrane capacitance from 37.2 (+/-7.3) to 27.4 (+/-6.1) mF/m2, and also showed that accompanying changes in the mean internal electrical conductivity and dielectric permittivity of the cells were insignificant. Scanning electron microscopy was used to examine the surface morphology of the cells and, in agreement with our previous reports for leukemia cells, the observed membrane capacitance values correlated closely with the morphological complexity of the cell membrane surface. The observed changes in the membrane dielectric properties are discussed in terms of their biological significance and their relationship to previously-detected changes in cell surface charge.

Animals↗

Sustained slow sinus rhythm entrained by retrograde atrial impulses.

Sustained slow sinus rhythm due to a returning cycle after a premature atrial P wave is described. It seems possible to consider a model of the forced vibration in a spring-mass system or resonant electric circuit of an alternating current series element containing resistance R, inductance L, and capacitance C as a quantitative approach to describing the slow sinus rhythm. The differential equation expressing cardiac resonant oscillation has for the case R = 500 omega the form 178 (d2X/dt2) + 500(dX/dt) + 5600X = 4490 cos 3.57t, where X is 100q, where q is the capacitor charge. By solving the equation of this special case, we obtained an effective voltage and current of 0.84 V and 1.68 mA, respectively. Determination of cardiac electrical activity by this method was considered to be useful, and may have practical applications in solving problems of nearly linear resonance in vital phenomena. This is the first paper on applying a linear second order differential equation to describing a frequency response modelling of slow sinus rhythm due to entrainment by the retrograde atrial impulses.

Aged↗

The onset and development of transmission in the chick ciliary ganglion.

1. The onset and development of transmission has been studied electro-physiologically in the isolated chick ciliary ganglion from Stage 25 (Hamburger & Hamilton, 1951) until 28 days after hatching. Ultrastructure of the synapses was concomitantly investigated.2. Synaptic transmission began at Stage 26(1/2) and was 100% in both cell groups, ciliary and choroid, by Stage 33. It was initially chemical until Stage 41 when effective electrical coupling first appeared in the ciliary population. The proportion of electrically transmitting synapses increased to 80% by 1-2 days post-hatching.3. Few morphological synapses were present at Stage 33(1/2) when all ganglion cells were transmitting. A scarcity of synaptic vesicles persisted until late in embryonic development when all ciliary cells possessed calyces. At hatching the calyces were filled with synaptic vesicles.4. Initial synaptic contacts were by fine terminal branches often on the intricate processes of early ganglion cells. Calyces formed from Stage 36(1/2) and there was a concomitant retraction of ganglion cell processes, so that by Stage 40 all ciliary cells had simple calyces. The calyx was a transitory structure, which from the first week post-hatching began to break up into a cluster of boutons.5. Chemical post-synaptic potentials (PSPs) were at Stage 40 long (30 x the membrane time constant) and further prolonged by eserine. By Stage 43, PSPs had become markedly shortened and were unaffected by eserine. No simple explanation can be offered for the changes in PSP time course and sensitivity to anticholinesterases during development.6. Intracellular records from Stage 40 ciliary cells, which all possess calyces, showed 1-2 mV amplitude, diphasic, fast decaying electrical coupling potentials (CPs). Later in development the CPs became 20-40 mV amplitude, more slowly decaying and monophasic. This seemed to be correlated with faster presynaptic conduction velocities and myelination of the cell soma. Such changes in CPs may reflect a shift from capacitative to more resistive coupling and point to several factors contributing in varying degrees to the electrical transmission.7. Presynaptic fibres innervating ciliary cells were from the start of lower threshold and faster conduction velocity than those innervating ciliary cells, as occurred in the adult. It is concluded that these preganglionic fibres were probably specified by the time transmission starts and that they selectively innervated the proper post-synaptic cells.

Animals↗

The electrical circuitry of an olfactory sensillum in Antheraea polyphemus.

The aim of this study was to obtain a better insight into the distribution of active and passive electrical properties of the membranes of al olfactory sensillum. Such a sensillum is composed of three accessory cells which envelope the somata of two receptor neurons which have long (300 micron) dendrites that are incorporated in cuticular protuberances (hairs). An electrical equivalent circuit of this organ is reconstructed which is based upon the response of sensilla to current transients, the shape of extracellularly recorded spikes, and the effects of lesions on these signals. A strong influence of membrane capacitances on the overall response and on the shape of spikes becomes apparent. From the combination of electrical and morphometric data it follows that the resistance of the apical membranes of the accessory cells in much higher than the resistance of their basolateral membranes. Neither in simultaneous tip and sidewall recordings nor in the hair (dendrite) amputation experiments could antidromic propagation of action potentials into the dendrite be demonstrated. The commonly accepted retrograde propagation explanation for the biphasic waveform of the spike is, therefore, rejected. Dendrite amputation experiments showed that the impedance of the dendrite does not limit the spike amplitude and that the dendrite contributes slightly to the duration of the positive phases of spikes. Both of these observations indicate that the dendrite is also involved in the generation of the leading phase of the spike. Consequently the proposition is made that action potentials are initiated at a dendritic location.

Animals↗

A glass/silicon composite intracortical electrode array.

A new manufacturing technique has been developed for creating silicon-based, penetrating electrode arrays intended for implantation into cerebral cortex. The arrays consist of a 4.2 mm x 4.2 mm glass/silicon composite base, from which project 100 silicon needle-type electrodes in a 10 x 10 array. Each needle is approximately 1,500 microns long, 80 microns in diameter at the base, and tapers to a sharp point at the metalized tip. The technique used to manufacture these arrays differs from our previous method in that a glass dielectric, rather than a p-n-p junction, provides electrical isolation between the individual electrodes in the array. The new electrode arrays exhibit superior electrical properties to those described previously. We have measured interelectrode impedances of at least 10(13) omega, and interelectrode capacitances of approximately 50 fF for the new arrays. In this paper, we describe the manufacturing techniques used to create the arrays, focusing on the dielectric isolation technique, and discuss the electrical and mechanical characteristics of these arrays.

Electric Conductivity↗

Extraction of all coefficients of coupled-mode equations for natural, single-phase, unidirectional SAW transducers from dispersion characteristics computed by hybrid finite element method.

A numerical method is presented for determining all of the coefficients of coupled-mode equations for natural, single-phase, unidirectional SAW transducers (NSP-UDTs). Substituting numerical results computed by the hybrid finite element method for infinite NSPUDTs with shorted and open electric ports into several relations derived from the coupled-mode theory, we can determine all of the coefficients. Specifically, the edge frequencies of a stop-band and the static capacitances yield the self-coupling coefficients and the amplitudes of mutual-coupling and transduction coefficients, and the electric potential standing wave on the substrate surface, which can be derived from the projection of the standing wave distributions of particle displacements and electric potential in the whole substrate onto the set of those predicted by the coupled-mode theory, yields the phases of mutual-coupling and transduction coefficients. NSPUDTs on ST-25 degrees X quartz, Y-51.25 degrees Z LiTaO3, and 50 degrees Y-25 degrees X La3Ga5SiO14 substrates are investigated. Our results agree well with the earlier experimental ones.

Journal Article↗

Influence of conductance changes on patch clamp capacitance measurements using a lock-in amplifier and limitations of the phase tracking technique.

We characterized the influence of conductance changes on whole-cell patch clamp capacitance measurements with a lock-in amplifier and the limitations of the phase-tracking method by numerical computer simulations, error formulas, and experimental tests. At correct phase setting, the artifacts in the capacitance measurement due to activation of linear conductances are small. The cross talk into the capacitance trace is well approximately by the second-order term in the Taylor expansion of the admittance. In the case of nonlinear current-voltage relationships, the measured conductance corresponds to the slope conductance in the range of the sine wave amplitude, and the cross talk into the capacitance trace corresponds to the second-order effect of the slope conductance. The finite gating kinetics of voltage-dependent channels generate phase-shifted currents. These lead to major artifacts in the capacitance measurements when the angular frequency of the sine wave is close to the kinetic rate constant of the channel. However, when the channel kinetics are sufficiently slow, or sufficiently fast, the cross talk is still close to the second-order effect of the measured conductance. The effects of activation of voltage-dependent currents on the capacitance measurements may be estimated, provided a detailed characterization of the kinetics and voltage dependence is available. A phase error of the lock-in amplifier of a few degrees leads to significant projections. The phase-tracking method can be used to keep the phase aligned only during periods of low membrane conductance. However, nonideal properties of the equivalent circuit, in particular the fast capacitance between the pipette and the bath solutions, may lead to large phase errors when the phase-tracking method is used, depending on the electrical properties of the cell. In this article we provide practical values, setting the range where possible artifacts are below defined limits. For proper evaluation of capacitance measurements, the capacitance and conductance traces should always be displayed together.

Animals↗

Electrophysiological characteristics of cultured human umbilical vein endothelial cells.

Electrophysiological characteristics of cultured human umbilical vein endothelial cells (HUVEC) were determined using the patch-clamp technique in the whole cell configuration. In isolated cells, membrane potential, capacitance, and input resistance were (Mean +/- SD) - 16.3 +/- 12.7 mV, 53.9 +/- 26 pF, and 2.3 +/- 1.3 G omega, respectively (N = 26); and in confluent cells - 23.6 +/- 5.5 mV, 127 +/- 59 pF, and 0.254 +/- 0.077 G omega, respectively (N = 6). The almost 10 times higher input resistance, and smaller capacitance of isolated versus confluent cells, indicated that the latter were in electrical communication, presumably through open gap junctions, which was confirmed by intercellular diffusion of Lucifer Yellow. Whole-cell currents of isolated cells were made up of at least three components: First, two outward currents, an early transient one with activation-inactivation kinetics and a small delayed sustained component with 6.75 +/- 4.8 and 0.73 +/- 0.089 nS conductance, respectively. Second, an inward component which was rectified and had 1.58 +/-1.2 nS conductance. In contrast to a reported lack of voltage-gated channels in HUVEC, the above currents were voltage dependent. Inhibition of the whole-cell currents by external Ba2, internal Cs, and other K+ blockers indicates that the three observed currents are carried by K+. This was confirmed by changes of outside K+ concentrations shifting the I-V curve intercept in the direction expected for K(+)-selective channels. Voltage-gated Ca2+ currents were not apparent in the whole-cell current records. HUVEC membrane potential was as low as that of microvascular cells, while inward current rectification at normal external K+ was like that in arterial endothelial cells. This mixed phenotypic expression and multipotential behavior suggests that the electrical features of HUVEC may be primarily determined by embryonic origin and the local effect of the microenvironment rather than strictly by vessel size.

Calcium Channels↗