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ISFET-neuron junction: circuit models and extracellular signal simulations.

Purpose of this paper is to characterize the Ion-Sensitive Field-Effect Transistors (ISFET)-neuron junction, based on the equivalent electric-circuit approach. As a result, recording of action potentials can be simulated with a general-purpose circuit simulation program such as HSPICE. The neuronal electrical activity, extracellularly recorded by the ISFET, is analyzed as a function of the physical-chemical and geometric ISFET parameters, of the ionic currents in the neuron, and of the neuro-electronic junction parameters such as the sealing resistance, double-layer capacitance, and general adhesion conditions. The models of the neuron, of the coupling circuit, and of the ISFET implemented in HSPICE are first described. These models are then used to simulate the behavior of the junction between a patch of neuronal membrane (described by the compartmental model) and the ISFET.

Biophysical Phenomena↗

Effects of double-layer polarization on ion transport.

It has been proposed that changes in ionic strength will alter the shape of current-voltage relations for ion transport across a lipid membrane. To investigate this effect, we measured currents across glyceryl monooleate membranes at applied potentials between 10 and 300 mV using either gramicidin and 1 mM NaCl or valinomycin and 1 mM KCl. A bridge circuit with an integrator as null detector was used to separate the capacitative and ionic components of the current. The changes in the current-voltage relations when ionic strength is varied between 1 and 100 mM are compared with predictions of Gouy-Chapman theory for the effects of these variations on polarization of the electrical diffuse double-layer. Double-layer polarization accounts adequately for the changes observed using membranes made permeable by either gramicidin or valinomycin.

Biological Transport↗

Mathematical model that characterizes transmitral and pulmonary venous flow velocity patterns.

The transmitral and pulmonary venous flow velocity (TMFV and PVFV, respectively) patterns are related to the physiological state of the left heart by use of an electrical analog model. Filling of left ventricle (LV) through the mitral valve is characterized by a quadratic Bernoulli's resistance in series with an inertance. Filling of the left atrium (LA) through the pulmonary veins is represented by a lumped network of linear resistance, capacitance, and inertance. LV and LA are each represented by a time-varying elastance. A volume dependency is incorporated into the LV model to produce physiological pressure-volume loops and Starling curves. The state-space representation of the analog model consists of 10 simultaneous differential equations, which are solved by numerical integration. Model validity is supported by the following. First, the expected effects of aging and decreasing LV compliance on TMFV and PVFV are accurately represented by the model. Second, the model-generated TMFV and PVFV waveforms fit well to pulsed-Doppler recordings in normal and postinfarct patients. It is shown that the TMFV deceleration time is prolonged by the increase in LV compliance and, to a lesser extent, by the increase in LA compliance. A shift from diastolic dominance to systolic dominance in PVFV occurs when LA compliance or pulmonary perfusion pressure increases or when LV compliance or mitral valve area decreases. The present model should serve as a useful theoretical basis for echocardiographic evaluation of LV and LA functions.

Animals↗

The collapse of the sarcoplasmic reticulum in skeletal muscle.

When various cations, including Ca2+, are in the fixative, both sarcoplasmic reticulum (SR) of whole skeletal muscle and isolated SR vesicles collapse to form pentalaminate "compound membranes" that result from the apparent fusion of the lumenal lamellae of the membranous envelope of the SR. The process may be reversed by subsequently soaking the tissue in 1 M NaCl. An identical morphological phenomenon is observed in unfixed quickly frozen isolated frog skeletal muscle fibers, the cation in that case coming from endogenous sources. The hypothesis is advanced that the collapse is an in vivo process mediated by the sequestration of Ca2+ after contraction. The resulting obliteration of the SR lumen would have the effect of displacing the SR contents into the junctional SR, as well as electrically isolating the free SR from the junctional SR during relaxation. As a consequence, resistive coupling between the plasmalemma and the junctional SR becomes a plausible mechanism for the translation of the action potential into Ca2+ release, since the bulk of the SR membrane capacitance would now remain separated from the plasmalemma during relaxation.

Animals↗

Elastic, dielectric, and piezoelectric losses in piezoceramics: how it works all together.

The quality factor along with electromechanical coupling coefficient (CEMC) is commonly used as a measure of the energy efficiency of a piezoelectric transducer (PT) working as an energy converter. Losses in piezoceramics are phenomenologically considered to have three coupled mechanisms: dielectric, elastic, and piezoelectric. Their cumulative performance first of all determines the PT quality factor characterizing the efficiency of vibrational energy accumulation, and related to it dissipative effects. The extended definition of the PT electromechanical quality factor (EMQ) with permanent energy exchange between electrical source of excitation and PT was proposed. The EMQ analysis has been conducted on the basis of complex material constants for both stiffened and unstiffened canonical vibrational modes. The efficiency of mechanically free and electrically excited piezoceramic transducers in a wide frequency range of PT harmonics, especially between the fundamental resonance and antiresonance frequencies, was investigated, and the effect of piezoelectric loss anomaly with extremely low total losses was predicted. Particularly, optimization of PT excitation with connected reactive (capacitive) element was conducted to provide higher PT mechanical vibrational characteristics with less total losses. The requirements to the piezoceramic material parameters, types of transducer vibrations, and especially to the piezoelectric loss factor in the range of physically valid values were established to provide maximal EMQ.

Journal Article↗

Relation of electrical properties of skin to structure and physiologic state.

Interpretation of cutaneous electrical measurements in terms of structure or function requires special techniques for identifying the separate contributions of the various elements. Resistance measures have different implications depending upon whether the subject is sweating and whether the electrode preparation is wet or dry. Microelectrode measurement indicates that the stratum corneum and other epidermal layers represent a significant pathway for ion conductance. Impedance measurement allows estimation of skin capacitance and of the thickness of the capacitative element. When applied to data on excised stratum corneum, this analysis indicates the presence of a relatively impermeable layer less than 2 microns thick, a conclusion subject to some doubt because of uncertainty over the dielectric constant of wet keratin. The use of impedance measurement with closely spaced electrodes gives an indication of the hydration of the superficial horny layer and also demonstrates reabsorption of sweat from this region. Potential measurement at the surface reflects the relative internal resistance of two parallel sources, the sweat glands and an "epidermal generator."

Electric Conductivity↗

Changes in cellular membrane and paracellular conductances by amphotericin B in the epithelium of the bullfrog cornea.

In the isolated bullfrog cornea, measurements of DC electrical parameters in conjunction with AC impedance and ultrastructural analyses were used to determine the effects of 10(-5) M amphotericin B on epithelial cellular membrane and paracellular conductances. In NaCl Ringers, amphotericin B elicited a 3.5-fold increase in the specific apical membrane conductance (Ga/Ca); where Ga and Ca are the apical membrane conductance and capacitance, respectively. The basolateral membrane conductance (Gb) and the basolateral membrane capacitance (Cb) fell by 57% and 50%, respectively. In the paracellular pathway, the tight junctional complex (Gj) was unchanged whereas the lateral intercellular space resistance (Rp) decreased by 55%. The declines in Gb and Cb were suggestive of cell volume shrinkage because these changes were consistent with a previously described decline in intracellular K+ content and reduction in exposed basolateral membrane area to current flow. Ultrastructural analysis validated that amphotericin B caused cell volume shrinkage because there was: (1) increased folding of the basolateral membrane and waviness of the basal aspects- of the plasma membrane; (2) dilatation of the lateral intercellular spaces. This agreement suggests that intracellular activity decreased following exposure to amphotericin B which resulted in cell volume shrinkage and an impairment of Cl- uptake across the basolateral membrane.

Amphotericin B↗

Electric cell-substrate impedance sensing (ECIS) as a noninvasive means to monitor the kinetics of cell spreading to artificial surfaces.

This article describes the optimization of an experimental technique referred to as electric cell-substrate impedance sensing (ECIS) to monitor attachment and spreading of mammalian cells quantitatively and in real time. The method is based on measuring changes in AC impedance of small gold-film electrodes deposited on a culture dish and used as growth substrate. Based on experimental data and theoretical considerations we demonstrate that high-frequency capacitance measurements (f = 40 kHz) are most suited to follow the increasing surface coverage of the electrode due to cell spreading. The excellent time resolution of the method allowed an in-depth analysis of cell spreading kinetics under various experimental conditions. Using ECIS we studied the attachment and spreading of epithelial MDCK cells (strain II) on different protein coatings, and investigated the influence of divalent cations on spreading kinetics. We quantified the inhibitory effect of soluble peptides that mimic the recognition sequence of fibronectin and other extracellular matrix proteins (RGDS). We also applied the ECIS technique to monitor the detachment of confluent fibroblastic cell layers (WI38/VA-13) by means of these peptides.

Animals↗

Morphological and electrophysiological properties of single myocardial cells from Koch triangle of rabbit heart.

BACKGROUND: The morphological and electrophysiological characteristics of cardiac cells in Koch triangle are still disputed. We studied the appearance and electrical properties of these diverse myocytes to elucidate their complex electrophysiological phenomena. METHODS: Experiments were conducted using cooled charge coupling device (CCD) system and whole cell, patch clamp technique to determine the morphology, action potential and sodium current density of single viable myocytes enzymatically isolated from the Koch triangle of rabbit hearts. RESULTS: Morphologically, cardiac cells in shape of spider, tiny spindle, slender spindle, rod and strip were observed in percentage of 3.0 +/- 0.3, 35.0 +/- 5.0, 15.0 +/- 2.0, 40.0 +/- 5.0 and 6.0 +/- 0.7 respectively. The cellular dimensions and capacitance gradually increased in the above order (all P < 0.05). Electrophysiologically, action potential configurations recorded from them were similar respectively to nodal (N), atrial nodal (AN), nodal Hisian (NH), atrial (A) and Hisian like potentials obtained from the intact atrioventricular nodal preparations. Diastolic depolarization appeared in all myocytes except for rod cells. Sodium current density increased in the order of tiny spindle, strip, rod, slender spindle cell (all P < 0.05), but could not be detected in spider-shaped cells. Linear regression analysis revealed that membrane capacitance was correlated negatively to the rate of diastolic depolarization r = -0.70, P < 0.001, but positively to maximum depolarization potential, amplitude of action potential, upstroke velocity and maximum peak value of sodium current density r = 0.84, 0.80, 0.87 and 0.75, respectively; all P < 0.001. CONCLUSIONS: The results demonstrated that spider-shaped, spindle, rod and strip cells in Koch triangle might correspond to pacemaking, transitional, atrial and Purkinje like cells, respectively. Furthermore, tiny spindle and slender spindle cells were referred to transitional cell alpha (TCalpha) and beta (TCbeta) accordingly considering their distinctive electrical properties. Different myocytes with diverse electrical properties constituted the infrastructure of sophisticated electrophysiological phenomena in Koch triangle. In view of the prominent percentage and electrical properties, tiny spindle and slender spindle cells were presumed to play important roles.

Action Potentials↗

Cation recognition by self-assembled monolayers of oriented helical peptides having a crown ether unit.

Cation recognition of self-assembled monolayers (SAMs) of helical peptides having a crown ether unit was investigated by the impedance spectroscopy and cyclic voltammetry. Lipo-(Ala-Aib)8-Ala-Cr and Boc-Glu(Cr)-(Ala-Aib)8-Lipoa (Lipo, Lipoa, and Cr represent lipoic acid, lipoamide, and amidobenzo-18-crown-6, respectively) were synthesized and the helix SAMs were prepared. The peptides having a crown ether unit formed SAMs oriented nearly vertically to the substrate. The capacitance of the Lipo-(Ala-Aib)(8)-Ala-Cr SAM changed specifically with the addition of cations, and the binding constants of the SAM were larger than those of the crown ether in aqueous solution because of a large dipole moment of the helical peptide. In the case of the Boc-Glu(Cr)-(Ala-Aib)8-Lipoa SAM, the cation binding to the SAM showed a drastic decrease in the peak current of the cyclic voltammetry around 10(-5)M of K+ ion. In either capacitance measurement or cyclic voltammetry, the helical peptide SAM played an important role in the sensitive response to cations.

Cations↗

Transport-dependent alterations of membrane properties of mammalian colon measured using impedance analysis.

Direct current (DC) measurement methods have been commonly used to characterize the conductance properties of the mammalian colon. However, these methods provide no information concerning the effects of tissue morphology on the electrophysiological properties of this epithelium. For example, distribution of membrane resistances along narrow fluid-filled spaces such as the lateral intercellular spaces (LIS) or colonic crypts can influence DC measurements of apical and basolateral membrane properties. We used impedance analysis to determine the extent of such distributed resistance effects and to assess the conductance and capacitance properties of the colon. Because capacitance is proportional to membrane area, this method provides new information concerning membrane areas and specific ionic conductances for these membranes. We measured transepithelial impedance under three conditions: control conditions in which the epithelium was open-circuited and bathed on both sides with NaCl-HCO3 Ringer's solutions, amiloride conditions which were similar to control except that 100 microM amiloride was present in the mucosal bathing solution, and mucosal NaCl-free conditions in which mucosal Na and Cl were replaced by potassium and sulfate or gluconate ("K+ Ringer's"). Three morphologically-based equivalent circuit models were used to evaluate the data: a lumped model (which ignores LIS resistance), a LIS distributed model (distributed basolateral membrane impedance) and a crypt-distributed model (distributed apical membrane impedance). To estimate membrane impedances, an independent measurement of paracellular conductance (Gs) was incorporated in the analysis. Although distributed models yielded improved fits of the data, the distributed and lumped models produced similar estimates of membrane parameters. The predicted effects of distributed resistances on DC microelectrode measurements were largest for the LIS-distributed model. LIS-distributed effects would cause a 12-15% underestimate of membrane resistance ratio (Ra/Rb) for the control and amiloride conditions and a 34% underestimate for the "K Ringer's" condition. Distributed resistance effects arising from the crypts would produce a 1-2% overestimate of Ra/Rb. Apical and basolateral membrane impedances differed in the three different experimental conditions. For control conditions, apical membrane capacitance averaged 21 microF/cm2 and the mean apical membrane specific conductance (Ga-norm) was 0.17 mS/microF. The average basolateral membrane capacitance was 11 microF/cm2 with a mean specific conductance (Gb-norm) of 1.27 mS/microF.(ABSTRACT TRUNCATED AT 400 WORDS)

Amiloride↗

Electrophysiology of the marine diatom Coscinodiscus wailesii IV: types of non-linear current-voltage-time relationships recorded with single saw-tooth voltage-clamp experiments.

Electrophysiological states of the marine diatom Coscinodiscus wailesii are known to change spontaneously in the temporal range of seconds. In order to assess the genuine current-voltage-time relationships of individual states in less than a second, voltage-clamp experiments have been carried out using single sweeps of saw-tooth shaped command voltages. This method is introduced with model calculations. Plotting the results in current-voltage coordinates provides convenient access to several electrophysiological entities, such as absence of drift (smoothly closed IV loops), membrane capacitance (by I jump at sign reversal of dV/dt), and ohmic conductances (in linear regions of the current-voltage relationship), as well as equilibrium voltage (internal intersection of capacitance-corrected, 8-shaped tracings) and coarse gating kinetics (rise or fall of capacitance-corrected I at sign reversal of dV/dt) of a voltage-sensitive ion conductance. From electrophysiological measurements with double-barreled glass-microelectrodes on C. wailesii, several distinct types of current-voltage loops are presented. Most of the data, including recordings from electrical excitation, can be interpreted as temporal relaxations of voltage-sensitive conductances for K(+) and Cl(-). A more detailed analysis of the effect of tetraethylammonium (TEA(+)) shows that 10 and 20 mM TEA(+) inhibit the K(+) conductance in C. wailesii only by up to about 20% but predominantly via a K(+) outward rectifier.

Diatoms↗

Changes in Friend murine erythroleukaemia cell membranes during induced differentiation determined by electrorotation.

We used electrorotation measurements to investigate alterations in the plasma membranes of DS19 murine erythroleukaemia cells that accompanied erythropoietic differentiation induced by hexamethylene bisacetamide (HMBA). Following 3 days of HMBA treatment, the mean cell membrane specific capacitance determined from electrorotation spectra of individual, viable cells at physiological tonicity (300 mosmol/kg) fell from 1.74 to 1.53 microF/cm2, in agreement with trends observed earlier by dielectrophoretic measurements on bulk cell populations. Scanning and transmission electron microscopy revealed that the relatively high values found for cell membrane capacitance (> 1 microF/cm2) reflected the large area of plasma membrane associated with complex surface morphology including numerous microvilli. Furthermore, it demonstrated that the fall in membrane capacitance during HMBA treatment correlated with a reduction in the density of these complex surface features. Differences in the mechanical characteristics of the cell membranes of untreated and treated cells were then examined by exposing cells to osmotic stress. The intricacy of membrane morphology intensified with increasing osmolality of the suspending medium and this was reflected in higher specific capacitance values. When the osmolality was increased from 210 to 450 mosmol/kg, the mean membrane capacitance of untreated DS19 cells changed from 1.58 to 2.05 microF/cm2 while that for HMBA-treated cells changed from 1.47 to 1.72 microF/cm2, a significantly smaller response. This demonstrated that cells exposed to 72 h of differentiation treatment had an enhanced mechanical resilience as compared with their untreated counterparts, evidencing the early stages of the development of the membrane skeleton which becomes fully developed in mature erythrocytes. Our findings demonstrate the value of electrorotation measurements as a method for the non-invasive characterisation of viable leukaemic cells and their responses to stimuli and show that the membrane capacitance values so derived reflect membrane morphology.

Acetamides↗

A quantitative analysis of the voltage-current relationships of fixed charge membranes and the associated property of "punch-through".

A theoretical analysis of the voltage-current relationship is carried out in a membrane consisting of two fixed charge regions, of opposite sign, in contact. This is achieved by applying the diffusion equations to this system in conjunction with the Poisson-Boltzmann equation. The latter has been successfully applied by Mauro to determine the profiles of the electrostatic potential in his treatment of the capacitative property of such a system. It is shown that the system displays the property of rectification and is very similar in many respects to a solid state P-N junction diode. It is also shown that for the case of reverse bias, an electrical breakdown phenomena can occur. This is referred to as the "punch-through" effect. "Punch-through" was observed in experiments on the electrical characteristics of the membranes of Chara australis and Nitella sp. The experimental results are discussed in relation to the theoretical analysis.

Cell Membrane↗

Capacitive monitoring of protein immobilization and antigen-antibody reactions on monomolecular alkylthiol films on gold electrodes.

Self-assembled monolayers of omega-mercaptohexadecanoic acid and omega-mercaptohexadecylamine on gold electrodes are stable at neutral pH and display pure capacitive behavior at frequencies around 20 Hz. Different methods of covalent immobilization of proteins on these monolayers are compared. Various reagents including succinimides, thionylchloride, p-nitrophenol and carbodiimides were used to activate the carboxy groups of the adsorbed monolayer of omega-mercaptohexadecanoic acid. Glutaraldehyde, cyanuric chloride and phenylene diisocyanate were used to activate the amino groups of the monolayer of omega-mercaptohexadecylamine. The immobilization of albumin on the activated surface was studied by capacitive measurements. The N-hydroxysuccinimide and carbodiimide methods were identified as most suitable for protein immobilization in that they did not compromise the insulating properties of the alkylthiol layer and led to maximal increase of its dielectric thickness. These approaches were used for a layer-by-layer preparation of a capacitive immunosensor. Specifically, antibodies to human serum albumin were immobilized on the alkylthiol mono-layer. Binding of the antigen led to a decrease of the electrode capacitance. The detection limit of the immunosensor is as low as 15 nM (1 mg/l).

Alkylation↗

Effects of spiperone on self-stimulation and other activities of the Mongolian gerbil.

1 Self-stimulation to lever pressing and capacitance probe touching was obtained in Mongolian gerbils (Meriones unguiculatus) from electrode placements within the medial forebrain bundle. 2 Lever pressing was more sensitive to the decremental effects of a central depressant, pentobarbitone, than capacitance probe touching, suggesting its greater responsiveness to disturbances of motor function. 3 Spiperone (0.005 to 0.05 mg/kg) attenuated capacitance probe touching and lever pressing equally, a finding explained by action on either reward pathways or on the ability to initiate responding. 4 This same dose range of spiperone (0.005 to 0.05 mg/kg) attenuated locomotor activity, whether spontaneous or evoked by non-contingent electrical stimulation, and produced catalepsy. 5 The spiperone-induced attentuation of self-stimulation was not necessarily a result of its action on dopaminergic reward pathways since the effects could equally well be explained by a failure to initiate responding.

Animals↗

Effect of sympathetic nerve stimulation and adrenoceptor blockade on pial arterial and venous calibre and on intracranial pressure in the cat.

Pial arterial and venous calibre were continuously recorded through a closed cranial window preparation during cervical sympathetic nerve stimulation in 10 cats before and after alpha- and beta-adrenoceptor blockade. In addition, the intracranial pressure (ICP) was simultaneously recorded in 4 of the cats. Under resting conditions 33 arteries (mean diameter 130 micron) constricted by 11.7 +/- 0.8% and 80 venous portions (mean diameter 152 micron) constricted by 13.7 +/- 0.7% during sympathetic nerve stimulation. ICP decreased simultaneously by 16.5 +/- 6.2%. Administration of the alpha-adrenoceptor antagonist phenoxybenzamine i.v. (1.5 mg X kg-1) abolished the reduction of ICP and markedly reduced, but did not completely abolish, the constrictor response of arteries and veins. The beta-adrenoceptor antagonist propranolol (1.5 mg X kg-1) did not significantly alter the reduction of ICP or the response of pial veins and small arteries, whereas the response of arteries with a diameter greater than 150 micron was attenuated. It is concluded that the constriction of pial veins and arteries during sympathetic stimulation is mediated predominantly via alpha-adrenoceptors. The sympathetic nerves of cerebral blood vessels may have stronger influence on the cerebral capacitance than on resistance vessels under normotensive and normocapnic conditions.

Adrenergic alpha-Antagonists↗

A neuron-silicon junction: a Retzius cell of the leech on an insulated-gate field-effect transistor.

An identified neuron of the leech, a Retzius cell, has been attached to the open gate of a p-channel field-effect transistor. Action potentials, spontaneous or stimulated, modulate directly the source-drain current in silicon. The electronic signals match the shape of the action potential. The average voltage on the gate was up to 25 percent of the intracellular voltage change. Occasionally weak signals that resemble the first derivative of the action potential were observed. The junctions can be described by a model that includes capacitive coupling of the plasma membrane and the gate oxide and that accounts for variable resistance of the seal.

Action Potentials↗