[Resistance and capacitance of Nitella during transient and constant conditions].
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Black lipid membranes made from oxidized cholesterol were examined for their specific resistance, capacitance, and physical stability, as a function of cholesterol oxidation time and of age. Membranes formed from cholesterol oxidized in n-octane were not physically stable even after 7 h of oxidation unless they were aged for at least a month. Membranes formed from cholesterol oxidized in decane and tetradecane (1 : 1) were stable immediately after 2--6 h of oxidation. Oxidation times outside this range produced unstable membranes. After 1 month storage, membranes from cholesterol solutions oxidized in decane and tetradecane from 0.75--3 h were stable. After 11 months, only the 0.75 oxidation time produced stable membranes. Storage in nitrogen retarded the aging process. After initial forming of the membrane, total membrane area and capacity increased and then stabilized, although specific capacity and resistance did not change, indicating inherent stability in the bilayer's intrinsic electrical properties. Bilayers formed soon after cholesterol oxidation had membrane capacity which ranged from 0.42 to 0.55 muF/cm2. Specific membrane resistance ranged initially from 2 . 10(6) to 37 . 10(6) omega/cm2 in 0.2 M NaCl with lower resistances in the more oxidized membranes. With aging, membrane capacity decreased gradually over 11 months to values approaching 0.1 muF/cm2 indicating membrane thickening. Membrane resistance ordinarily decreases with storage time. The rate of these changes with age is dependent on the extent of initial cholesterol oxidation and subsequent oxidation, with long term stability best in the least oxidized membranes.
This work combines a theoretical study of electrical interactions between two excitable heart cells, using a variable coupling resistance, with experimental studies on isolated rabbit ventricular cells coupled with a variable coupling resistance to a passive resistance and capacitance circuit. The theoretical results show that the response of an isolated cell to an increased frequency of stimulation is strongly altered by the presence of a coupling resistance to another cell. As the coupling resistance gradually is decreased, the stimulated cell becomes able to respond successfully to more rapid stimulation, and then, at levels of coupling resistance that allow conduction between the two cells, the coupled pair of cells exhibits arrhythmic interactions not predicted by the intrinsic properties of either cell. The experimental results show that the isolated rabbit ventricular cell is extremely sensitive to even a very small electrical load, with shortening of the action potential by 50% with electrical coupling to a model cell (of similar input resistance and capacitance to the ventricular cell) as high as 1,000 M omega, even though the action potential amplitude and current threshold are very insensitive to the electrical load.
Osteonecrosis of the femoral head in the adult is a progressive condition that, if untreated, usually results in femoral head collapse and secondary osteoarthritis. The experimental application of electrical and electromagnetic fields has been shown to favorably affect a number of biological processes pertinent to osteonecrosis of the femoral head and has led to several clinical trials. The condition has been treated by the application of electrical fields invasively by the surgical implantation of electrodes within the femoral head and noninvasively by capacitative or inductive coupling. This review describes results in osteonecrosis of the femoral head with these therapeutic techniques. Stimulation by means of inductive coupling with pulsed magnetic fields seems to be the most promising technique studied so far, but the optimal signal characteristics and device design are not yet known.
The electrical and morphological properties of human erythrocytes under high hydrostatic pressure up to 500 MPa have been studied by dielectric spectroscopy. The pressure-induced changes in the dielectric behavior of erythrocyte suspensions indicate that hydrostatic pressure causes the change in cell shape from discoidal to spherical and hemolysis at 200-300 MPa, the formation of buds and spicular processes followed by vesiculation at 300-400 MPa, and the increase in the membrane capacitance at 400-500 MPa.
These studies describe the effects of nanosecond (10-300 ns) pulsed electric fields (nsPEF) on mammalian cell structure and function. As the pulse durations decrease, effects on the plasma membrane (PM) decrease and effects on intracellular signal transduction mechanisms increase. When nsPEF-induced PM electroporation effects occur, they are distinct from classical PM electroporation effects, suggesting unique, nsPEF-induced PM modulations. In HL-60 cells, nsPEF that are well below the threshold for PM electroporation and apoptosis induction induce effects that are similar to purinergic agonistmediated calcium release from intracellular stores, which secondarily initiate capacitive calcium influx through store-operated calcium channels in the PM. NsPEF with durations and electric field intensities that do or do not cause PM electroporation, induce apoptosis in mammalian cells with a well-characterized phenotype typified by externalization of phosphatidylserine on the outer PM and activation of caspase proteases. Treatment of mouse fibrosarcoma tumors with nsPEF also results in apoptosis induction. When Jurkat cells were transfected by electroporation and then treated with nsPEF, green fluorescent protein expression was enhanced compared to electroporation alone. The results indicate that nsPEF activate intracellular mechanisms that can determine cell function and fate, providing an important new tool for probing signal transduction mechanisms that modulate cell structure and function and for potential therapeutic applications for cancer and gene therapy.
1. Whole-cell and perforated-patch tight-seal recording techniques were used to characterize the voltage-dependent membrane conductances of the Y-79 cells, a human retinoblastoma line composed of pluripotential retinal precursor cells. 2. Membrane resistance and capacitance were measured under current clamp, yielding approximate average values of 1.8 G omega and 26 pF, respectively. The cells are electrically excitable, and depolarization above -20 mV triggers slow action potentials. 3. Step depolarization of the membrane under voltage clamp elicits a high-threshold transient inward current, followed by a sustained, larger outward current. The outward current is carried by potassium ions, as determined by its susceptibility to blockage by K-channel antagonists [tetraethylammonium (TEA), Cs, and 4-aminopyridine (4-AP)] and insensitivity to reduction of external chloride concentration. 4. The isolated inward current displayed some unusual properties: its amplitude is directly related to extracellular calcium concentration, and replacement of calcium by magnesium completely abolishes it. However, none of the calcium channel antagonists tested (cadmium, nickel, nifedipine, and amiloride) exerted a substantial blockage. In addition, removal of external sodium or superfusion with tetrodotoxin significantly reduce the size of this current. 5. A single voltage-dependent conductance appears to underlie the inward current, because a variety of manipulations, such as changes in the holding potential, in the extracellular concentration of calcium or sodium, or superfusion with tetrodotoxin, failed to reveal the presence of kinetically distinct components. 6. The results suggest that a single voltage-dependent conductance mechanism underlies the depolarization-activated inward current in Y-79 cells. This channel appears to be primarily permeable to calcium, but with a significant contribution by sodium ions. Its functioning appears to be modulated by extracellular calcium.
Electrical measurements were carried out on planar lipid membranes from thylakoid lipids. The specific capacitance of membranes formed from decane-containing monogalactosyldiacylglycerol (MGDG), which accounts for 57% of the total lipid content of thylakoids, showed that it adopted a bilayer structure. Solvent-free bilayers of MGDG were not formed, with very rare exceptions, indicating that decane is required to stabilize the planar conformation. However, this cone-shaped lipid produces bilayer structures in combination with other cylindrical thylakoid lipids even in the absence of organic solvent. We compared the properties of solvent-free and decane-containing bilayers from MGDG, soybean lecithin, and the quaternary mixture of lipids similar to that found in vivo. The conductance of decane-MGDG was 26 times higher than that of decane-lecithin. The flux through the decane-lecithin bilayer was found to be slightly dependent on pH, whereas the decane-MGDG membrane was not. The specific conductance of bilayers formed from the quaternary mixture of lipids was 5 to 10 times larger than lecithin (with alkane or not). Further experiments with bilayers made in the presence of a KCl gradient showed that decane-MGDG, decane-MGDG/DGDG/SQDG/PG, and solvent-free MGDG/DGDG/SQDG/PG were cation-selective. The permeability coefficient for potassium ranged from 4.9 to 8.3 x 10(-11) cm s-1. The permeability coefficient for protons in galactolipids, however, was determined to be about six orders of magnitude higher than the value for potassium ions. The HCl permeation mechanism through the lipid membranes was determined from diffusion potentials measured in HCl gradients. Our results suggest that HCl was not transported as neutral molecules. The data is discussed with regard to the function of galactolipids in the ion transport through thylakoid membranes.
Transfer of the fluorescent dye, Lucifer yellow (LY), from an intracellularly injected neuron to one or more other neurons is accepted as indirect evidence of electrotonic interactions among such dye coupled cells. Direct evidence requires that at least two coupled cells be recorded from simultaneously and such evidence in the CNS has been gained only for hippocampal pyramidal neurons. Since interpretations of the functional significance of dye coupling among magnocellular neuroendocrine cells depend upon its relation to electrical coupling, we sought to obtain direct evidence for electrotonic interactions in such neurons. Over 150 pairs of supraoptic nucleus (SON) neurons in hypothalamic slices were recorded from intracellularly using one LY and one potassium acetate electrode in each instance. Of these, 9 pairs were studied in sufficient detail to determine that they were electrically coupled. Most of the remaining pairs were determined not to be coupled. In each coupled pair of cells, membrane voltage changes due to spontaneously occurring or current evoked action potentials, as well as current evoked hyperpolarizations, in one cell were reflected in similar, though attenuated changes in the other cell. All of these changes occurred simultaneously in the two neurons. Spontaneously arising postsynaptic potentials in the two cells were temporally uncorrelated. In each case that electrical coupling was observed, dye coupling resulted from LY injection. Coupling ratios ranged from 0.05 to 0.2. Capacitative coupling between the recording electrodes as an artifact was ruled out since cells in the same tissue penetration as the coupled cell showed no responses to membrane voltage changes in the primary cell; no responses were seen with the second electrode placed extracellularly or in the medium; and similar coupling potentials were also seen when one cell was recorded without a second electrode present. We conclude that electrical coupling exists among magnocellular neurons of the SON and that the incidence of dye coupling is a reasonable estimate of the incidence of electrical coupling. These electrotonic interactions probably play important roles in the coordination of firing among magnocellular neurosecretory neurons.
The tubular ascidian heart is composed of a single layer of cells joined together by apical (zonulae occludentes) and spot (maculae occludentes) nexuses. Intercalated discs or desmosomes were not observed in this tissue. Rectangular pulses of current were applied across the opened and flattened myocardium. Assuming that all the transepithelial current flowed through a uniform gap between cells, the resistivity in the gap must be very high compared to that in bulk solution. It is likely, therefore, that the gap width is of the order of an ionic radius or smaller. Assuming that all the transepithelial current flowed through the cells and that the inner and outer membranes had the same resistivity, the membrane resistivity was about 210 ohms cm(2) and the membrane capacitance was about 1.6 microF per cm(2). The myocardial cells were found to be in electrical continuity with each other through the nexuses since current could be passed through a strip of myocardium in a sucrose gap. Assuming that the longitudinal resistance of the cytoplasm was negligible, the cell-to-cell resistivity of the nexuses was 0.2 ohm cm(2). It is concluded that the nexuses provide a low resistance pathway between cells and a transepithelial barrier.
During the last decennia many protein-related electrical phenomena have been studied and applied in a variety of measuring systems, from simple metal electrodes with adsorbed proteins to sophisticated systems with lipid bilayers. Many of the investigations concern the monitoring of immuno reactions. The basic underlying electrical effects of the observed phenomena are the protein modulated dielectric constant, conductivity, electrical potential, ion permeability and ion mobility. In this paper special attention is paid to the capacitive measurements with EIS systems as well as impedance and potential measurements with FET devices. The Donnan theory is treated and applied to the static ImmunoFET operation, explaining the relatively small effects which have been reported. Finally, an alternative approach is described in which the ImmunoFET is applied in a dynamic way, to circumvent the drawbacks of the static measurements.
The adsorption of fullerols (C60(OH)n) on glutathione-adsorbed gold electrode was characterized by using double-impedance system, i.e., electrochemical quartz crystal impedance and electrochemical impedance spectroscopy, and cyclic voltammetry. The time courses of piezoelectric parameters were used to reflect the changes of interfacial physical properties, such as mass, density-viscosity, and dielectric constant, during the adsorptions of peptide and fullerols onto electrode. The electrochemical impedance based on the simple equivalent electric network were also simultaneously measured and provided electrochemical interface information, e.g., double-layer capacitance and charge-transfer resistance. It was found that the double-impedance responses were varied with the forms of glutathione. It was also shown that the frequency curves due to the adsorption of oxidized (GSSG) and reduced (GSH) glutathione could be exhibited as different kinetic equations. The heterogeneous charge-transfer rate constants of ferricyanide/ferrocyanide before and after the peptide and fullerols adsorption were determined by CV and EIS methods. The results showed that the proposed method has potential applications in interfacial studies of biomaterials, since these combined techniques have advantages in real time providing multidimensional piezoelectric and electrochemical impedance information.
Using voltage-time-dependent negative resistance characteristics of Voltage-Current curves of excitable cell membranes estimated without using artificial voltage clamp method, the author made a quantitative analysis of excitability of cell membranes and different conditions of transmembrane action potentials as a bias voltage to the negative resistance of the excitable cell membrane. The pacemaker cells were classified as "Astable Oscillators" and nonpacemaker excitable cells as "Monostable Oscillators," and application of a rapidly changing electromagnetic field to the cells was analyzed as a means of stimulating the cells. The understanding of the 10 essential electrical parameters is highly desirable for safe and effective electrical stimulation. Among these, emphasis was placed on the often neglected, important electrical parameters of "output impedance" of stimulation pulse wave complexes for + and - polarity components, as well as the importance of capacitive current (Ic = C.dV/dt) which depends on rise time as well as fall time of the stimulation pulse wave, and undesirable side effects of electrolysis phenomena due to excessive D.C. current. The difference and similarity between TENS (Transcutaneous Nerve Stimulation) and TES (Transcutaneous Electrical Stimulation), TENMS (Transcutaneous Electrical Nerve and Muscle Stimulation) or TMS (Transcutaneous Muscle Stimulation) was discussed. The author's clinical study indicated that effective TES (TENMS)--characterized by effective muscle contraction without creating pain with a pulse repetition rate approximately the same as the heart rate of the individual--can often give superior beneficial effects in improvement of micro-circulation and subsequent relief of pain and other symptoms compared with TENS that creates stimulation of large diameter sensory nerve fibers without creating significant muscle contraction. Such improvement is often accompanied by the abolishment of the pain with disappearance of local substance P and increase in local serotonin with disappearance of local L-tryptophan.
The proteinoid cells are assembled of thermal polymers of amino acids. Typically, an appropriate mixture of amino acids containing aspartic or glutamic acid is heated at 190 degrees C for 6 h, stirred with water for 2 h, dialyzed during 2 d, and lyophilized. Spheroidal cells are made from such polymer by dissolving it in the water by boiling, and then cooling. Many of them can be made by sonication at room temperature. These artificial cells, ranging from microns to tens of microns in diameter (depending on composition and preparation), have double membranes and various internal compositions. The spherules can microencapsulate dyes, oxidant-reductant compounds or acceptor-donor substances, and can be packed together. Such spherules display electrical polarization and electrical discharges and respond to intra- and extracellular ionic and electric influence upon membrane and action potential. These properties arise from the double membrane structure, asymmetric membrane permeability, and channeling phenomena. Such features as exponential dependence of the steady-state conductance and capacitance as well as negative resistance of the membrane seem to be responsible for the flip-flop alternations of the membrane polarization, rhythmic electric oscillations, and all-or-none action potentials. The presence of such chromophores as pteridine and flavin in polymers constituting these cells is responsible for their photosensitivity.
Present instruments used in pulsed field and conventional gel electrophoresis, encounter a number of real and conceived difficulties such as electrical hazard, ground leak current, electrical noises, formation of gas bubbles at the metal electrodes and production of Joule heat in the buffer-gel system. To overcome the above-mentioned problems a novel electrophoresis unit based on capacitively-induced pulsed field was designed and tested in which the applied high voltage is decoupled from the electrolyte (buffer-gel system). In order to achieve a higher performance, the primary pulse generator which has been fabricated to apply for capacitively-induced pulsed field electrophoresis was equipped with a modulating frequency pulse generator which produced a combination of low and high frequency waves. The newly designed electrophoresis unit was able to resolve the Lambda DNA fragments so that seven bands with an acceptable resolution were observed. By increasing the run time both the depletion of molecules from the wells and the resolution of the bands improved compare to the patterns obtained via conventional electrophoresis.
The systolic hump in the aortic blood pressure wave is defined as the aortic-resistance component proportional to the aortic blood flow superimposed on the windkessel component. An electrical analogue comprising a series resistance (aortic resistance) plus a resistance (peripheral resistance) and capacitance (aortic compliance) in parallel (i.e. windkessel component) is used for analysis. Curve fitting using the least-squares method is performed on calculated and measured blood pressure waves from dogs under haemodynamical conditions induced by infusion of three drugs (noradrenaline, isoproterenol and acetylcholine). The curve fitting RMS (root mean square) errors are < 3% for blood pressure waves and < 30% for blood flow waves, with good agreement between measured and calculated blood flow waveforms. Infusion of noradrenaline and acetylcholine is found to induce a significant decrease and increase in the aortic resistance, respectively. Although only a small fraction of the blood pressure wave, the systolic hump has a marked effect on the systolic pressure waveform.
Adrenal medullary chromaffin cells release catecholamines and neuropeptides in an activity-dependent manner controlled by the sympathetic nervous system. Under basal sympathetic tone, catecholamines are preferentially secreted. During acute stress, increased sympathetic firing evokes release of both catecholamines as well as neuropeptides. Both signalling molecules are co-packaged in the same large dense core granules, thus release of neuropeptide transmitters must be regulated after granule fusion with the cell surface. Previous work has indicated this may be achieved through a size-exclusion mechanism whereby, under basal sympathetic firing, the catecholamines are selectively released through a restricted fusion pore, while less-soluble neuropeptides are left behind in the dense core. Only under the elevated firing experienced during the sympathetic stress response do the granules fully collapse to expel catecholamines and neuropeptides. However, mechanistic description and physiological regulation of this process remain to be determined. We employ electrochemical amperometry, fluid-phase dye uptake and electrophysiological capacitance noise analysis to probe the fusion intermediate in mouse chromaffin cells under physiological electrical stimulation. We show that basal firing rates result in the selective release of catecholamines through an Omega-form 'kiss and run' fusion event characterized by a narrow fusion pore. Increased firing raises calcium levels and activates protein kinase C, which then promotes fusion pore dilation until full granule collapse occurs. Our results demonstrate that the transition between 'kiss and run' and 'full collapse' exocytosis serves a vital physiological regulation in neuroendocrine chromaffin cells and help effect a proper acute stress response.
Capacitative Ca2+ entry (CCE) is a universal mechanism for refilling intracellular Ca2+ stores in electrically non-excitable cells. The situation in excitable cells is less clear, however, since they may rely on other entry mechanisms for Ca2+-store refilling. In the present study we investigated CCE in intact PC12 cells, using acetylcholine to bring about activation of InsP3 receptors (InsP3Rs), caffeine to activate ryanodine receptors (RyRs) and thapsigargin to inhibit sarco/endoplasmic reticulum Ca2+-ATPase pumps. We found that depletion of the InsP3-, caffeine- or thapsigargin-sensitive stores promoted Ca2+ entry, suggesting that stimulation of either InsP3Rs or RyRs can activate CCE. The CCE pathways activated by InsP3Rs, RyRs and thapsigargin appeared to be independent at least in part, since their effects were found to be additive. However, CCE triggered by caffeine, acetylcholine or thapsigargin progressively diminished with time. The decay of CCE caused by one agent also inhibited subsequent responses to the others, suggesting that some component of the CCE pathway is common to all intracellular Ca2+ stores. The magnitude of CCE stimulated by InsP3Rs or RyRs was related to the size of the stores; the InsP3-sensitive store was smaller than the RyR-sensitive store and triggered a smaller entry component. However, both stores filled with a similar half time (about 1 min), and both could be filled more rapidly by depolarization-induced Ca2+ entry through voltage-operated channels. A significant basal Ca2+ influx was apparent in PC12 cells. The basal entry component may be under the control of the InsP3-sensitive Ca2+ store, since short incubations in Ca2+-free medium depleted this store.