[Dynamic capacitance electrode for the takeoff of bioelectrical potentials].
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Electric Conductivity↗
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Transient and permanent fusion of vesicles in Zea mays coleoptile protoplasts measured in the cell-attached configuration.
Exocytosis in protoplasts from Zea mays L. coleoptiles was studied using patch-clamp techniques. Fusion of individual vesicles with the plasma membrane was monitored as a step increase of the membrane capacitance (Cm). Vesicle fusion was observed as (i) An irreversible step increase in Cm. (ii) Occasionally, irreversible Cm steps were preceded by transient changes in Cm, suggesting that the electrical connection between the vesicle with the plasma membrane opens and closes reversibly before full connection is achieved. (iii) Most frequently, however, stepwise transient changes in Cm did not lead to an irreversible Cm step. Within one patch of membrane capacitance steps due to transient and irreversible fusions were of similar amplitude. This suggests that the exocytosis events do not result from the fusion of vesicles with different sizes but are due to kinetically different states in a fusion process of the same vesicle type. The dwell time histogram of the transient fusion events peaked at about 100 msec. Fusion can be described with a circular three-state model for the fusion process of two fused states and one nonfused state. It predicts that energy input is required to drive the system into a prevailing direction.
Cell Membrane↗
Electrical properties of rabbit corneal endothelium as determined from impedance measurements.
Alternating- and direct-current electrical characteristics of rabbit corneal endothelium were studied under varying experimental conditions. The measurements were performed by sending a 10-microA current (AC or DC) across the tissue layer. Maximal values of transendothelial potential difference and resistance were 1.3 +/- 0.1 mV and 73 +/- 6 omega . cm2, respectively. The short-circuit current was estimated from the potential and resistance values. Impedance loci were obtained for the frequency range 0.5-100 kHz. A capacitive reactance (C = 0.63 +/- 0.02 microF/cm2) was observed in the 100 Hz-100 kHz range. To relate the impedance data to the electrical parameters of the cell membranes, the voltage-divider ratio was determined by sending square pulse across the tissue and measuring voltage responses across the apical and basal membranes with an intracellular microelectrode. The intracellular potential difference was on the average -61 +/- 1 mV, and the voltage-divider ratio was found to be between 0.33 and 4. Impedance data were fit by a computer to an equivalent circuit representing a "lumped" model, and the agreement between the model and the data was satisfactory. The results are discussed in terms of both the morphological characteristics and properties of the fluid transport mechanism across the preparation.
Animals↗
The selection of electromyographic needle electrodes.
Monopolar and concentric reusable and disposable EMG needle electrodes from several manufacturers were studied at various stages of usage. Needle tips were examined and measured microscopically, and needles were tested for spontaneous noise and resistive and capacitative characteristics at four different test signal frequencies. Resistive and capacitative characteristics were found to be frequency dependent. Needle tip areas differed by as much as 10 times among the various electrodes. The amount of noise correlated most highly with resistive characteristics. Monopolar and concentric needles differed in respect to tip area, noise, and all resistive and capacitative measures. Coaxial disposable and reusable electrodes did not differ significantly and minor differences were noted in electrical characteristics between reusable and disposable monopolar needles, even from the same manufacturer. There was considerable variability among manufacturers for both needle types. In choosing an electrode, consistent tip area is probably the most important consideration to ensure repeatable quantifiable results, but minimizing needle impedance is useful to reduce noise.
Artifacts↗
Recovering the passive properties of tapered dendrites from single and dual potential recordings.
We demonstrate that measurement of the membrane potential at one or more sites on a branched and tapered neuron following a known transient injection of subthreshold somatic current uniquely determines the cell's passive electrical properties. That is, knowledge of the potentials allows recovery of the cell's axial resistance, membrane capacitance, membrane conductance and soma conductance. The argument underlying uniqueness leads immediately to a constructive, robust algorithm that we successfully test on synthetic data. The robustness stems from the fact that the algorithm requires only a few weighted integrals, or moments, of the measured potentials.
Algorithms↗
Synaptic- and agonist-induced excitatory currents of Purkinje cells in rat cerebellar slices.
1. Postsynaptic currents originating from activation of the two major excitatory inputs to Purkinje cells were studied in thin slices of rat cerebellum, using the tight-seal whole-cell recording technique. Two types of excitatory postsynaptic currents were analysed: those evoked by stimulation of the granule cell-parallel fibre system (PF-EPSC) and those elicited by stimulation of the climbing fibres (CF-EPSC). 2. Both types of postsynaptic currents had a linear current-voltage relation, reversing at membrane potentials close to 0 mV. Their time course of activation was independent of the membrane potential. 3. For both types of postsynaptic currents, the time course of decay was well described by a single exponential function, with a time constant which increased as the membrane potential was made more positive. 4. Postsynaptic currents arising from stimulation of the climbing fibre generally had a slightly faster time course of onset and decay than those associated with stimulation of the granule cell-parallel fibre system. The average values of the 10-90% rise time were 1.8 +/- 0.4 ms (means +/- S.D., n = 7) for PF-EPSCs and 0.8 +/- 0.3 ms (n = 9) for CF-EPSCs. Time constants of decay, at a holding potential of -60 mV, had values of 8.3 +/- 1.6 ms (n = 7) and 6.4 +/- 1.1 ms (n = 9) for PF-EPSCs and CF-EPSCs respectively. 5. CF-EPSCs and PF-EPSCs had the characteristics described above in slices derived from animals aged 9-22 days old and 9-15 days old, respectively. The PF-EPSCs in animals older than 15 days had very slow time courses and positive apparent reversal potentials, suggesting that they originated from distal locations, not under accurate voltage control. 6. In order to assess the quality of the voltage clamp, responses to hyperpolarizing pulses from -70 mV were analysed. The capacitive currents could be fitted by the sum of two exponentials, and were interpreted with an equivalent electrical circuit comprising two main compartments (soma and proximal dendrites on one hand, distal dendrites on the other). Analysis of synaptic currents in terms of this model suggested that the recorded time course of decay was approximately correct. 7. CF-EPSCs as well as PF-EPSCs were insensitive to the NMDA receptor antagonist 3-3(2-carboxypiperazine-4-yl)propyl-1-phosphonate (CPP), but were blocked in a dose-dependent reversible manner by the non-NMDA antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX).(ABSTRACT TRUNCATED AT 400 WORDS)
6-Cyano-7-nitroquinoxaline-2,3-dione↗
Extracellular ATP activates calcium signaling, ion, and fluid transport in retinal pigment epithelium.
The presence of receptors for ATP has not been established in any native preparation of retinal neurons or glia. In the present study, we used conventional electrophysiological and [Ca2+]in fluorescence imaging techniques to investigate the effects of ATP added to Ringer's solution perfusing the retinal-facing (apical) membrane of freshly isolated monolayers of bovine retinal pigment epithelium (RPE). ATP (or UTP) produced large, biphasic voltage and resistance changes with a Kd of approximately 5 microM for ATP and approximately 1 microM for UTP. Electrical and pharmacological evidence indicates that the first and second phases of the response are attributable to an increase in basolateral membrane Cl conductance and a decrease in apical membrane K conductance, respectively. The ATP-induced responses were not affected by adenosine, but were reduced by the P2-purinoceptor blocker suramin. ATP also produced a large, transient increase in [Ca2+]in that was blocked by cyclopiazonic acid, an inhibitor of endoplasmic reticulum Ca2+-ATPases. The calcium buffer BAPTA attenuated the voltage effects of ATP. We also found that apical DIDS significantly inhibited the ATP-evoked [Ca2+]in and electrical responses, suggesting that DIDS blocked the purinoceptor. Measurements of fluid movement across the RPE using the capacitance probe technique demonstrated a significant increase in fluid absorption by apical UTP. These data indicate the presence of metabotropic P2Y/P2U-purinoceptors at the RPE apical membrane and implicate extracellular ATP in vivo as a retinal signaling molecule that could help regulate the hydration and chemical composition of the subretinal space.
4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
Perpetual atrial arrhythmias causing a prolonged returning cycle as an expression of cardiac resonant oscillation.
A theoretical method of calculating ventricular myocardial voltage and time constant from the findings of 40 portions in electrocardiographic (ECG) tracings of 25 cases, in which atrial premature beats with full returning cycle can be observed, is presented. An resonant electric circuit of an alternating current series element containing resistance (R) of 500 ohms, inductance (L), and capacitance (C) was assumed as a model in this study. The steady-state voltage (Vm) was calculated in terms of the real part of the complex expression L(d2q/dt2) + 500(dq/dt) + (1/C)q = Vmej omega t, where L = 250/ zeta omega n, 1/C = omega n2 L, and Vm = A(zero) omega n2 Lx 100. The effective voltage derived ranged from 0.77 to 0.88 (mean, 0.82) V. The mean value approximated the mean voltage stimulation threshould of the human right ventricle. The time constant RC ranged from 73 to 135 ms. This is the first paper to establish that this method is clinically applicable. The dynamical system regulating the heart rhythm was described as a finite-dimensional matrix representation, which possesses diagonal elements, i.e. eigenvalues, of I/4 square root of 2 pi and 1/2.
Arrhythmias, Cardiac↗
Changes in electrophysiological properties of rat skin with age.
The age-related changes in the electrical and physiological properties of the skin were examined in rats at the ages of 5, 10, 21, 90, and 180 d. The resistance of the stratum corneum, the resistance of the viable skin (epidermis and dermis), and the capacitance of the stratum corneum were analyzed from skin impedance data using an equivalent circuit. With development and aging, the resistance of the stratum corneum and the viable skin increased, whereas the capacitance of the stratum corneum decreased. Physiological characteristics such as the thickness of skin strata and the content of lipid and water in the stratum corneum were also measured. The lipid content in the stratum corneum was constant at all ages. The water content in the stratum corneum decreased, and the thickness of skin strata increased with age. Comparison between electrical data and physiological properties suggested that the increase in the resistance of the stratum corneum with aging is primarily caused by the decrease in the water content and that the capacitance of the stratum corneum and the resistance of the viable skin depend on age-related increases in the thickness of skin strata. In conclusion, the age dependency of cutaneous electrical properties may affect the permeation profile of drugs through the skin, and impedance analysis can be used to estimate age-related changes in transdermal drug delivery.
Animals↗
Effect of intracellular pH on depolarization-evoked calcium influx in human sperm.
Human sperm are endowed with putative voltage-dependent calcium channels (VDCC) that produce measurable increases in intracellular calcium concentration ([Ca(2+)](i)) in response to membrane depolarization with potassium. These channels are blocked by nickel, inactivate in 1-2 min in calcium-deprived medium, and are remarkably stimulated by NH(4)Cl, suggesting a role for intracellular pH (pH(i)). In a previous work, we showed that calcium permeability through these channels increases approximately onefold during in vitro "capacitation," a calcium-dependent process that sperm require to fertilize eggs. In this work, we have determined the pH(i) dependence of sperm VDCC. Simultaneous depolarization and pH(i) alkalinization with NH(4)Cl induced an [Ca(2+)](i) increase that depended on the amount of NH(4)Cl added. VDCC stimulation as a function of pH(i) showed a sigmoid curve in the 6.6-7.2 pH(i) range, with a half-maximum stimulation at pH approximately 7.00. At higher pH(i) (> or =7.3), a further stimulation occurred. Calcium release from internal stores did not contribute to the stimulating effect of pH(i) because the [Ca(2+)](i) increase induced by progesterone, which opens a calcium permeability pathway that does not involve gating of VDCC, was unaffected by ammonium. The ratio of pH(i)-stimulated-to-nonstimulated calcium influx was nearly constant at different test depolarization values. Likewise, depolarization-induced calcium influx in pH(i)-stimulated and nonstimulated cells was equally blocked by nickel. In our capacitating conditions pH(i) increased 0.11 pH units, suggesting that the calcium influx stimulation observed during sperm capacitation might be partially caused by pH(i) alkalinization. Additionally, a calcium permeability pathway triggered exclusively by pH(i) alkalinization was detected.
Ammonium Chloride↗
Maturation of synapses and electrical properties of cells in the cochlear nuclei.
Auditory nerve fibers carry impulses from the cochlea to the cochlear nuclei. There the temporal firing patterns of auditory nerve fibers are preserved by some cells and altered by others. The two factors which govern how firing patterns are shaped are (1) the intrinsic electrical properties of cells that determine the size and time course of voltage changes caused by synaptic currents and (2) the synaptic circuitry between cells. The electrical properties of cells were measured by recording the responses to current injected intracellularly into brain slice preparations. The synaptic responses to electrical shocks of the auditory nerve were used to determine the functional properties of synaptic connections. The three distinct types of electrical properties of cells that can be distinguished electrophysiologically in similar preparations of mature tissue, bushy and stellate cells in the ventral cochlear nucleus [(1984) J. Neurosci. 4, 1577-1588] and cells in the dorsal cochlear nucleus [Hirsch and Oertel (1987) (submitted); Oertel et al. (1987) In: Functions of the Auditory System, Editor: S. Hassler. J. Wiley and Sons (in press)] can be differentiated at least as early as 7 days after birth. Young cells, however, have higher input resistances and lower input capacitances than mature cells, and they cannot sustain high firing rates. Bushy and stellate cells in the ventral cochlear nucleus respond to electrical stimulation of the auditory nerve with both excitatory and inhibitory postsynaptic potentials as early as 4 days after birth. The synaptic potentials occur with longer and more variable latencies than in mature cells and synapses fatigue more easily, however. Cells of the dorsal cochlear nucleus also receive both excitatory and inhibitory synaptic inputs 4 days after birth, upon stimulation of the auditory nerve. No systematic changes were detected in these synaptic responses as a function of age but this may have been because the variability in the shape and timing of synaptic responses was large even in mature tissue.
Action Potentials↗
Transient charge accumulation in a capacitive self-assembled monolayer.
Charge accumulation in an organosilane monolayer self-assembled on silicon is studied using electron-spectroscopy-based chemically resolved electrical measurements (CREM). By resolving the net electrical response of the organic layer, a significant capability of holding extra charge is indicated. Quantum size effects at a molecularly thin layer and the role of competing discharge mechanisms, including defect-assisted leakage currents, are discussed.
Electric Conductivity↗
An isolation power supply by phototransmission.
An isolation power supply by phototransmission was proposed, and the conversion efficiency and the electric characteristics of a prototype of the power supply unit were estimated. Infrared light emitting diodes (LEDs) and single-silicon-crystal solar cells were used as light sources and photovoltaic cells, respectively. A maximum conversion efficiency of 3.9% was achieved. The constant output voltage feedback system, which detects the change of the output voltage of the solar cells and controls the driving current of the LEDs, operates this power supply at maximum conversion efficiency independent of loads. The liquid-cooling structure of the power supply unit can keep the conversion efficiency nearly constant independent of the input power. The fabricated power supply has little capacitive leakage current and negligibly small internal noise.
Electric Power Supplies↗
[Electrical characteristics of the sensory neurons of the medical leech].
The electrical characteristics of three types of sensory neurons (N, T and P) were determined in the leech Hirudo medicinalis. Statistically significant differences were found between the values of tau, Cs and Rs in the neurons of N- and T-types. The same parameters show the least variation coefficients. The value of time constant (tau) is 17.000-46 ms, the resistivity of the soma membrane TRs) is 19.50 kphi-cm2 in T-neurons and 35 kphi-cm2 in N-neurons. The surface of the neurons estimated by their capacitance (Cs) can be 4-10 times larger than the surface of a sphere of the same size. The possible influence of electrical characteristics of the neurons on the peculiarities of their electrical reactions is discussed.
Action Potentials↗
Electrical properties of isolated demyelinated rat nerve fibres.
Isolated large rat nerve fibres with diphtheria toxin induced paranodal demyelination were investigated. These fibres had increased membrane time constant of the demyelinated nodal segment, related to a large increase in capacitance (5 to 50 times). The resting conductance was less increased (2 to 3 times), meaning that the internodal axolemma has considerably higher resting resistance than the nodal membrane. A large variation in action potential amplitude was found which was unrelated to the size of the nodal widening and the passive electrical properties.
Action Potentials↗
Use of AC impedance analysis to study membrane changes related to acid secretion in amphibian gastric mucosa.
We have applied transepithelial AC impedance techniques to gastric mucosa to reconcile ultrastructural and electrophysiological findings about gastric acid secretion and the mucosal barrier. By fitting impedance data measured at different HCl secretion rates to equivalent circuit models, we extracted capacitances and resistances (as measures of membrane area and ionic conductance, respectively) for the apical and basolateral membranes. The impedance measurements were found to be incompatible with earlier equivalent circuit models that modeled membrane electrical properties as lumped circuits based on one or two cell types. A distributed circuit model was developed that assumed only one dominant electrical pathway (i.e., one cell type), but that incorporated electrical effects arising from long and narrow membrane-lined structures present in the epithelium (e.g., gastric crypts, tubulovesicles, lateral intercellular spaces). This morphologically based model was found to represent the measured data accurately, and to yield values for membrane capacitances consistent with morphometric measurements of membrane areas. The main physiological conclusions from this analysis were as follows: (a) The dominant transepithelial current pathway may reside in the oxyntic cells. (b) The transepithelial conductance increase associated with the onset of acid secretion is entirely due to increased conductance of the apical membrane. This is in turn due entirely to increased area of this membrane, resulting from incorporation of tubulovesicular membrane. (c) When membrane conductances are normalized to actual membrane area by use of membrane capacitances, it turns out that acid secretion is not associated with a change in specific ionic conductance (change in conductance per unit area) at either the apical or basolateral membrane. (d) The puzzlingly low value of transepithelial resistance (</=400 Omega-cm(2)) arises because there are hundreds or thousands of square centimeters of actual membrane area per square centimeter chamber area. Apical membrane resistance is 25 kOmega-cm(2) (actual membrane area), implying a tight barrier to back-diffusion of protons.
Animals↗
Two-microelectrode voltage clamp of Xenopus oocytes: voltage errors and compensation for local current flow.
Oocytes from Xenopus laevis are commonly used as an expression system for ion channel proteins. The most common method for their electrophysiological investigation is the two-microelectrode voltage clamp technique. The quality of voltage clamp recordings obtained with this technique is poor when membrane currents are large and when rapid charging of the membrane is desired. Detailed mathematical modeling of the experimental setup shows that the reasons for this weak performance are the electrical properties of the oocytes and the geometry of the setup. We measured the cytosolic conductivity to be approximately 5 times lower than that of the typical bath solution, and the specific membrane capacitance to be approximately 6 times higher than that of a simple lipid bilayer. The diameter of oocytes is typically approximately 1 mm, whereas the penetration depth of the microelectrodes is limited to approximately 100 microm. This eccentric current injection, in combination with the large time constants caused by the low conductivity and the high capacitance, yields large deviations from isopotentiality that decay slowly with time constants of up to 150 micros. The inhomogeneity of the membrane potential can be greatly reduced by introducing an additional, extracellular current-passing electrode. The geometrical and electrical parameters of the setup are optimized and initial experiments show that this method should allow for faster and more uniform control of membrane potential.
Animals↗
Membrane admittance of cloned muscle cells in culture: use of a micropipet technique.
The impedance characteristics of muscle cells have been investigated by many investigators because of its unique features. The studies of the electrical properties of cloned cultured muscle cells, however, are scant and only a handful of papers have been published. In this report, the admittance of cultured Myoblast L6 cells was investigated. The capacitance and conductance of mononucleate myoblast cell membrane were found to be 2.2-2.5 microF/cm2 and 50-60 microS/cm2 and independent of frequency. The capacitance of 2.2 microF/cm2 is larger than the widely accepted capacitance value for most of the biological cell membranes, i.e., 1 microF/cm2. In contrast, those found for fused multinucleate myotubes are 5-6 microF/cm2 and 0.4 mS/cm2 and exhibited a marked frequency dependence. We postulate that the large and frequency dependent capacitance of myotubes does not arise from the surface membrane alone but is due to a tubule-like structure which may be similar to those found in frog muscles. Analysis of the results using an equivalent circuit demonstrates that the capacitance of myotubes is independent of frequency when the correction is made for the series resistances which are distributed in the lumen of tubule-like structure. An alternative explanation for the frequency dependent capacitance is based on the use of the transmission line theory or cable equation. The cable theory predicts that measured capacitance and conductance are inherently a function of frequency regardless of the nature of membrane capacitance and conductance if the dimension of cells is larger than or comparable to the propagation constant.
Animals↗