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PubMed1997

A theoretical basis for the high-frequency performance of the outer hair cell's receptor potential.

The frequency response of the outer hair cell (OHC) was studied theoretically. An electrical model of the OHC was analyzed mathematically, taking into account the effect of its inherent voltage-dependent capacitance. It was found that the variations of the capacitance dependent on the membrane potential could enhance the high-frequency response of the OHC, so that its cutoff frequency could be extended into the audio range. It was found further that the enhancement of the frequency response of the OHC was strongly dependent on its resting potential and on the ratio of the maximum voltage-dependent capacitance to the membrane linear capacitance.

Hair Cells, Auditory↗
PubMed1975

Analog circuit of the Acetabularia membrane.

The high membrane potential of Acetabularia (Em=-170mV) is due to an electrogenic pump in parallel with the passive diffusion system (Ed=-80mV) which could be studied separately in the cold, when the pump is blocked. Electrical measurements under normal conditions show that the pump pathway consists of its electromotive force Ep with two elements P1 and P2 in series; P2 is shunted by a large capacitance (Cp=3mF cm-2). The nonlinear current-voltage relationship of P1 (light- and temperature-sensitive) could be determined separately; it reflects the properties of a carrier-mediated electrogenic pump. The value of Ep(-190 mV) indicates a stoichiometry of 2:1 between electrogenically transported charges and ATP. The electrical energy normally stored in Cp, compares well with the metabolic energy, stored in the ATP pool. The nonlinear current-voltage relationship of P2 (attributed to phosphorylating reactions) is also sensitive to light and temperature and is responsible for the region of negative conductance of the overall current-voltage relationship. The power of the pump (1 muW cm-2) amounts to some percent of the total energy turnover. The high Cl- fluxes (1 nmol cm-2 sec-1) and the electrical properties of the plasmalemma are not as closely related as assumed previously. For kinetic reasons, a direct and specific Cl- pathway between the vacuole and outside is postulated to exist.

Acetabularia↗
PubMed1985

Depolarizing afterpotentials in myelinated axons of mammalian spinal cord.

Microelectrode recordings were made from 5-10 micron dia axons of adult rat spinal cord in vitro. Action potentials in response to electrical stimulation were recorded intracellularly and electrical characteristics of the axons were examined by injecting current pulses through a bridge circuit. All action potentials larger in amplitude than 80 mV were followed by depolarizing afterpotentials, similar to those recorded in peripheral axons [Barrett and Barrett (1982) J. Physiol., Lond. 323, 117-144]. The afterpotential could be described as the sum of three exponential components, the time constants of which (tau 1, tau 2 and tau 3) were 25.2 +/- 5.6, 3.1 +/- 0.8 and 0.8 +/0 0.3 ms, respectively, at 25 degrees C and a membrane potential of -80 mV. The maximal amplitudes of the afterpotential components, obtained by extrapolating to the peak of the action potential, were 3.8 +/- 1.0, 6.4 +/- 5.2 and 21.7 +/- 9.8 mV, for action potential amplitudes of 102 +/- 11 mV. The amplitude of the longest component of the afterpotential decreased with depolarization and increased with hyperpolarization at the recording site. The amplitude decreased markedly with increase of temperature to physiological levels, in conjunction with the expected decrease in action potential duration. Similar afterpotential components were present in the response of the axon to injected hyperpolarizing current pulses. The observations are consistent with the suggestion [Barrett and Barrett (1982) J. Physiol., Lond. 323, 117-144] that the afterpotential results from charging of the axolemmal capacitance by current passing through the myelin sheath during the action potential. They are inconsistent with a number of calculations of electrical characteristics of peripheral axons derived from voltage clamp experiments in isolated fibers. It is argued that the electrical resistance of the myelin lamellae is relatively low, though within the range calculated for other glial membranes. This suggestion is found more compatible with the available morphological data than the alternative proposal that a leakage pathway under the myelin sheath might be responsible for the afterpotential [Barrett and Barrett (1982) J. Physiol., Lond. 323, 117-144]. The significance of this organization for the function of myelinated axons and the electrical basis of the afterpotential are examined further in the accompanying paper [Blight (1985) Neuroscience 15, 13-31].

Action Potentials↗
PubMed1994

DNA, protein, and plasma-membrane incorporation by arrested mammalian cells.

Incorporation of DNA, protein, and plasma membrane during blockage by aphidicolin or by doxorubicin was studied by flow cytometry and electrorotation of three cell lines (mouse-myeloma Sp2/0-Ag14, hybridoma H73C11, and fibroblast-like L929 cells). Drug-mediated arrest at the G1-S boundary (aphidicolin) or in G2/M (doxorubicin) did not arrest synthesis of either protein or total membrane area, the increases in which outstripped growth in cell volume and apparent cell area, respectively. Measurements of membrane capacity in normal and hypo-osmotic media showed that the drugs had not changed the fundamental bilayer, but that an increase in the number or size of microvilli must have occurred. Aphidicolin-arrested cells withstood hypo-osmotic stress better than untreated cells could, indicating that the membrane excess can be utilized as a reserve during rapid cell expansion. Hypo-osmotically treated cell populations exhibited only about half the coefficient of variance (CV) in membrane properties of cells at physiological osmolality. Populations of arrested cells exhibited the same high CV as asynchronous cells, indicating that chemical arrest does not give uniformly villated cell populations. However, the lowest CV values were given by some synchronized (aphidicolin-blocked, then released) populations. Removal of aphidicolin allowed most cells to progress through S and G2, and then divide. During these processes, the membrane excess was reduced. After removal of doxorubicin, the cells did not divide: some continued protein synthesis, grew abnormally large, and further increased their membrane excess. Membrane breakdown by electric pulsing (3 x 5kV/cm, 40 microseconds decay time) of aphidicolin-synchronized L cells in G2/M led to a 22% loss of plasma membrane (both the area-specific and the whole-cell capacitance were reduced), presumably via endocytosis-like vesiculation.

Animals↗
PubMed2003

Membrane-bound and releasable nucleotidase activities: differences in canine mesenteric artery and vein.

1. At least two enzymatic activities are proposed to degrade the extracellular ATP: (i) ubiquitously expressed membrane-bound enzymes (ecto-nucleotidases); and (ii) soluble (releasable) nucleotidases that are released during stimulation of sympathetic nerves and break down neuronal ATP. No quantitative data have placed the magnitude of these nucleotidase activities into a physiological perspective of neurovascular control. 2. We studied comparatively the membrane-bound and releasable nucleotidase activities in canine isolated inferior mesenteric arteries and veins using 1,N6-etheno(epsilon)-nucleotides (i.e. epsilon-ATP, epsilon-ADP, epsilon-AMP and epsilon-adenosine) as exogenous substrates. The enzymatic activities were estimated by measuring the disappearance of the epsilon-substrate and appearance of epsilon-products by means of HPLC-fluorescence detection during either stimulation of sympathetic perivascular nerves (releasable activity) or in the absence of nerve stimulation (ecto-nucleotidase activity). 3. Incubation of vascular segments with 50 nmol/L epsilon-ATP for 60 min resulted in a decrease of the epsilon-ATP substrate by 63.5 +/- 4.6 and 91.2 +/- 6.2% in the artery and vein, respectively. In contrast, the decrease of the epsilon-ATP during electrical field stimulation (EFS; 16 Hz, 0.3 msec, 2 min) was 39.8 +/- 4.2% in the artery and 13.1 +/- 7.3% in the vein. Therefore, the mesenteric arteries demonstrate a greater releasable ATPase activity and a weaker ecto-ATPase activity than mesenteric veins. 4. The degradation of epsilon-ADP and epsilon-AMP was similar in both blood vessels under either experimental protocol. The epsilon-adenosine was not significantly degraded in the absence or presence of EFS. 5. These data implicate a differential removal of extracellular ATP as a potential mechanism of serving resistance and capacitance in the splanchnic circulation.

Animals↗
PubMed2005

[Measuring the electricity frequency properties of blood].

In order to understand the electricity frequency specialties of blood, we have developed a wide frequency electricity characteristic testing system and used it to test the amplitude frequency property and phase frequency property of the blood in different states and constituents at 1 Hz to 20 MHz. Further analysis on the results of tests helped us know some important properties of blood at even more microcosmic levels from a new angle. Meanwhile, some problems and considerations on the improvement of the electricity model of biotic tissue and blood were pointed out. (1) From 1 Hz to 5 KHz, the impedance of blood descended 99%. (2) Simple equivalent circuit of resistance and capacitance must be used in series equivalent but not in usual parallel connection equivalent. (3) Experiment indicated, equivalent circuits of blood need more analysis, because simple equivalent circuit of resistance and capacitance is liable to gross error. (4) When the three element model is used for measuring the resistance of inside liquid, capacitance of cell membrane and the resistance of outside liquid of blood, the three testing frequencies must be very similar.

Blood↗
PubMed2001

Inwardly rectifying K(+) channels in spermatogenic cells: functional expression and implication in sperm capacitation.

To fertilize, mammalian sperm must complete a maturational process called capacitation. It is thought that the membrane potential of sperm hyperpolarizes during capacitation, possibly due to the opening of K(+) channels, but electrophysiological evidence is lacking. In this report, using patch-clamp recordings obtained from isolated mouse spermatogenic cells we document the presence of a novel K(+)-selective inwardly rectifying current. Macroscopic current activated at membrane potentials below the equilibrium potential for K(+) and its magnitude was dependent on the external K(+) concentration. The channels selected K(+) over other monovalent cations. Current was virtually absent when external K(+) was replaced with Na(+) or N-methyl-D-glucamine. Addition of Cs(+) or Ba(2+) (IC(50) of approximately 15 microM) to the external solution effectively blocked K(+) current. Dialyzing the cells with a Mg(2+)-free solution did not affect channel activity. Cytosolic acidification reversibly inhibited the current. We verified that the resting membrane potential of mouse sperm changed from -52 +/- 6 to -66 +/- 9 mV during capacitation in vitro. Notably, application of 0.3-1 mM Ba(2+) during capacitation prevented this hyperpolarization and decreased the subsequent exocytotic response to zona pellucida. A mechanism is proposed whereby opening of inwardly rectifying K(+) channels may produce hyperpolarization under physiological conditions and contribute to the cellular changes that give rise to the capacitated state in mature sperm.

Animals↗
PubMed1992

Transmucosal impedance of small intestine: correlation with transport of sugars and amino acids.

Transmucosal impedances of isolated perfused segments of jejunum from mice and hamsters were measured at frequencies from 10-100,000 Hz in the presence and absence of sugars and amino acids. Na-coupled transport of organic substrates caused large decreases of transmucosal impedance, reflecting contraction of cytoskeletal proteins controlling permeability of tight junctions, functional surface of basolateral membranes, and width of extracellular pathways. The observed changes of impedance were closely correlated with molar rates of Na-coupled active transport rather than with molecular species. Thus amino acids and sugars having the same molar rates of active transport also have the same effects on transmucosal impedance. It is proposed that a nonspecific increase of intracellular osmotic pressure during active transport is the first step initiating cytoskeletal contraction. Cell volume regulatory responses, including increased basolateral K+ conductance and Ca2+ influx, may be subsequent steps leading to contraction of perijunctional actomyosin, formation of junctional dilatations, and exposure of lateral membranes. Enhancement of oxygen capacity of perfusion fluids (e.g., with fluorocarbon emulsion) is required to maintain viability of isolated intestinal epithelium; in plain oxygenated Ringer-HCO3 solution, the transmucosal impedance is abnormally low and cytoskeletal contractile responses to Na-coupled transport are attenuated. An electrical circuit analog is presented that simulates almost exactly the observed transmucosal impedances and provides quantitative evaluation of the effects of Na-coupled transport of sugars and amino acids on resistances of tight junctions, capacitance of basolateral membranes, and postjunctional resistances of lateral intercellular spaces and villus cores.

Amino Acids↗
PubMed1994

Analysis of decaying unitary currents in on-cell patches of cells with a high membrane resistance.

Under nonideal voltage-clamp conditions, unitary currents in the cell-attached patch (CAP) configuration of the patch-clamp technique are decaying instead of rectangular. From these data, four parameters can be determined when multiple identical channels are present. We analyzed an electrical equivalent circuit of the CAP configuration. This analysis yielded, for single-channel resistance, attached-cell membrane resistance, patch resistance, and attached-cell membrane capacitance, a set of different equations to calculate their values from these four parameters. To choose the most robust equation, those that yielded the same resistor/capacitance were compared on the basis of their sensitivity for inevitable deviations of the parameters from their nominal values. The results of our theoretical study are of methodological interest: 1) because they confirm that, under special conditions, CAP measurements can be used to measure electrical membrane properties of intact cells (this is important inasmuch as the CAP is the only configuration in which the cytoplasm of cells remains undisturbed) and 2) because we identify the level of voltage clamp and the equations with which the electrical properties can be determined with optimal accuracy.

Animals↗