Subthreshold sodium current from rapidly inactivating sodium channels drives spontaneous firing of tuberomammillary neurons

A role for "persistent," subthreshold, TTX-sensitive sodium current in driving the pacemaking of many central neurons has been proposed, but this has been impossible to test pharmacologically. Using isolated tuberomammillary neurons, we assessed the role of subthreshold sodium current in pacemaking by performing voltage-clamp experiments using a cell's own pacemaking cycle as voltage command. TTX-sensitive sodium current flows throughout the pacemaking cycle, even at voltages as negative as -70 mV, and this current is sufficient to drive spontaneous firing. When sodium channels underlying transient current were driven into slow inactivation by rapid stimulation, persistent current decreased in parallel, suggesting that persistent sodium current originates from subthreshold gating of the same sodium channels that underlie the phasic sodium current. This behavior of sodium channels may endow all neurons with an intrinsic propensity for rhythmic, spontaneous firing.     

A voltage-activated proton current in human cardiac fibroblasts

A voltage-activated proton current in human cardiac fibroblasts, measured using the whole-cell recording configuration of the patch-clamp technique, is reported. Increasing the pH of the bathing solution shifted the current activation threshold to more negative potentials and increased both the current amplitude and its rate of activation. Changing the pH gradient by one unit caused a 51mV shift in the reversal potential of the current, demonstrating a high selectivity for protons of the channel carrying the current. Extracellularly applied Zn(2+) reversibly inhibited the current. Activation of the current contributes to the resting membrane conductance under conditions of intracellular acidosis. It is proposed that this current in cardiac fibroblasts is involved in the regulation of the intracellular pH and the membrane potential under physiological conditions as well as in response to pathological conditions such as ischemia.     

Application of a pulsed electric field to cross-flow ultrafiltration of protein solution

The application of pulsed electric field was investigated in the crossflow ultrafiltration of BSA (bovine serum albumn) to economize the application time of electric current as well as to avoid inherent problems of long-term application of electric field. During the application of various cyclic patterns of pulsed electric current, the averaged filtration flowrate and the degree of concentration were maintained higher than those obtained in the absence of electric current application. The temperature increase, pH change, and BSA loss by electrodeposition were all negligible during the operation. The averaged filtration flowrate increased as the ON/OFF duration ratio of electric current was higher and as the period of ON/OFF cycle was shorter. The re-establishment of concentration polarization was dependent to the duration of current OFF state and, therefore, a longer duration of OFF state was not favorable in maintaining higher filtration flow rate. Although the averaged filtration flowrate was enhanced as the magnitude of electric current increased, the flowrate enhancement became smaller as the magnitude of current increased because there exists a current value above which the degree of electrokinetic depolarization is no further improved.     

Energy of Plasma Flows Accelerated in a Current Sheet as a Function of the Current Flowing through the Sheet

The energy of directed plasma flows accelerated in a current sheet is studied experimentally as a function of the current flowing through the sheet. It is found that the plasma flow energy rapidly increases with increasing current amplitude, nearly according to a power law with an exponent of 1.8–1.9. Using relations for the neutral current sheet and the concept of plasma acceleration under action of the Ampère forces, the energy of directed plasma flows is estimated as a function of the total current flowing through the sheet. It is shown that, as the current rises, the ion energy grows due to an increase in both the Ampère forces and the width of the current sheet within which plasma is accelerated.     

Measurement of electric current evoked by substrate transport via bi-directional H+/oligopeptide transporter over-expressed in HeLa cells: electrogenic efflux and existence of a newly observed channel-like state

In the present study, we measured an electric current induced by substrate transport in a HeLa cell over-expressing a human intestinal di/tri-peptide transporter using the whole-cell patch-clamp technique. Gly-Sar, a typical substrate, induced an inward current associated with its uptake, which showed concentration-dependency following Michaelis-Menten-type kinetics with an apparent K(0.5) of 1.3mM as well as voltage-dependency. An outward current accompanying the efflux of Gly-Sar was also observed after washing out the cell. This outward current was voltage-dependent and was reduced by the inward proton gradient. In the case of hydrophobic dipeptides such as Gly-Phe and Gly-Leu, a distinctive current was observed: after washing out the cells, no outward current was observed, but rather, an 'inward leak' current was sustained in spite of the absence of transportable substrate. This leaky current was abolished by the perfusion of Gly-Sar and subsequent washing. It is considered that the hydrophobic substrate sticks within the substrate-binding site and causes the newly observed state, or the 'inward leak' current.     

The Ventral Photoreceptor Cells of Limulus : III. A voltage-clamp study

In the dark, the ventral photoreceptor of Limulus exhibits time-variant currents under voltage-clamp conditions; that is, if the membrane potential of the cell is clamped to a depolarized value there is an initial large outward current which slowly declines to a steady level. The current-voltage relation of the cell in the dark is nonlinear. The only ion tested which has any effect on the current-voltage relation is potassium; high potassium shifts the reversal potential towards zero and introduces a negative slope-conductance region. When the cell is illuminated under voltage-clamp conditions, an additional current, the light-induced current, flows across the cell membrane. The time course of this current mimics the time course of the light response (receptor potential) in the unclamped cell; namely, an initial transient phase is followed by a steady-state phase. The amplitude of the peak transient current can be as large as 60 times the amplitude of the steady-state current, while in the unclamped cell the amplitude of the peak transient voltage never exceeds 4 times the amplitude of the steady-state voltage. The current-voltage relations of the additional light-induced current obtained for different instants of time are also nonlinear, but differ from the current-voltage relations of the dark current. The ions tested which have the greatest effect on the light-induced current are sodium and calcium; low sodium decreases the current, while low calcium increases the current. The data strongly support the hypothesis that two systems of electric current exist in the membrane. Thus the total ionic current which flows in the membrane is accounted for as the sum of a dark current and a light-induced current.     

Prevention of Pseudomonas aeruginosa adhesion by electric currents

The process of controlling bacterial adhesion using an electric current deserves attention because of its ease of automation and environmentally friendly nature. This study investigated the role of electric currents (negative, positive, alternating) for preventing adhesion of Pseudomonas aeruginosa and achieving bacterial inactivation. Indium tin oxide (ITO) film was used as a working electrode to observe adhesion and inactivation under electric polarization. Electric current types were classified into negative, positive, and alternating current. The working electrode acted as a cathode or anode by applying a negative or positive current, and an alternating current indicates that the negative current was combined sequentially with the positive current. The numbers of adhered cells were compared under a flow condition, and the in situ behavior of the bacterial cells and the extent of their inactivation were also investigated using time-lapse recording and live/dead staining, respectively. The application of a negative current prevented bacterial adhesion significantly (～81% at 15.0 μA cm(-2)). The positive current did not significantly inhibit adhesion (<20% at 15.0 μA cm(-2)), compared to the nonpolarized case. The alternating current had a similar effect as the negative current on preventing bacterial adhesion, but it also exhibited bactericidal effects, making it the most suitable method for bacterial adhesion control.     

Ion transport across the frog olfactory mucosa: the basal and odorant-stimulated states

The Ussing method was adapted to study the basal electrolyte transfer as well as the events that occur upon odorant stimulation in frog olfactory mucosa. The unstimulated short-circuit current was due mainly to a furosemide-sensitive ion transport system on the apical side of the olfactory mucosa. This current was not amiloride sensitive. The current-voltage relationship of the unstimulated state was linear. That of the odorant-evoked current was non-linear and amiloride-sensitive. Ouabain caused collapse of both the unstimulated and odorant-stimulated short-circuit current. In this case, voltage-clamping the tissue to non-zero values restored the odorant-evoked current with polarity depending on that of the clamping voltage. This suggested that the direction of the current is determined by that of the sodium electrochemical potential difference. Our results indicate that the unstimulated short-circuit current occurs through an apical sodium cotransport system, while the odorant-evoked current is due to odorant-activated, passive sodium channels that are amiloride sensitive.     

K+ and Ca++ currents in hair cells isolated from the semicircular canals of the frog]

Hair cells of the inner ear are endowed with different types of ionic channels. To characterize voltage- and ion-dependent channels in vestibular hair cells, experiments were performed in enzymatically isolated hair cells of frog semicircular canals by using the whole-cell configuration of the patch-clamp technique. A large outward current, identified as a K+ current, was recorded when 132 mM KCl were present in the pipette filling solution. It could be dissected pharmacologically into three different components. The first component, which was transient and selectively blocked by 10 mM external 4AP, is most likely an IA-type current. The second one, sensitive to 20 mM external TEA, might be a delayed rectifier K+ current, while the third component insensitive to TEA and showing faster activation time course has been interpreted as a K+ current of IKCa-type. After blocking the outward current by substituting Cs+ for K+ and adding 20 mM TEA to the internal solution, a sustained inward current, identified as a Ca++ current, could be recorded. This current did not inactivate, and was blocked by Cd++ more effectively than Ni++, thus suggesting the presence of Ca++ channels similar to the neuronal "L" channels. Since both K+ and Ca++ channels were recruited at potentials near the resting level, it is suggested that they are involved in the modulation of the resting as well as the evoked transmitter release from the basal pole of the hair cells.     

Large electrical currents traverse growing pollen tubes

Using a newly developed vibrating electrode, we have explored the electric fields around lily pollen germinating in vitro. From these field measurements, we infer that each weeted pollen drives a steady current of a few hundred picoamperes through itself. Considered as a flow of positive ions, this current enters an ungerminated grain's prospective growth site and leaves it opposite end. After a grain germinates and forms a tube, this current enters most of the growing tube and leaves the whole grain. The current densities over both of these extended surface regions are relatively uniform, and the boundary zone, near the tube's base, is relatively narrow. This current continues as long as the tube grows, and even continues when elongation, as well as cytoplasmic streaming, are blocked by 1 mug/ml of cytochalasin B. After a otherwise indistinguishable minority of tubes have grown to lengths of a millimeter or more, their current comes to include an endless train of discrete and characteristic current pulses as well as a steady component. These pulses are about 30s long, never overlap, recur every 60-100s, and seem to enter a region more restricted to be growing tip than the steady current's sink. In most ways, the current through growing lily pollen resembles that known to flow through focoid eggs.     

Prevention of Pseudomonas aeruginosa adhesion by electric currents

The process of controlling bacterial adhesion using an electric current deserves attention because of its ease of automation and environmentally friendly nature. This study investigated the role of electric currents (negative, positive, alternating) for preventing adhesion of Pseudomonas aeruginosa and achieving bacterial inactivation. Indium tin oxide (ITO) film was used as a working electrode to observe adhesion and inactivation under electric polarization. Electric current types were classified into negative, positive, and alternating current. The working electrode acted as a cathode or anode by applying a negative or positive current, and an alternating current indicates that the negative current was combined sequentially with the positive current. The numbers of adhered cells were compared under a flow condition, and the in situ behavior of the bacterial cells and the extent of their inactivation were also investigated using time-lapse recording and live/dead staining, respectively. The application of a negative current prevented bacterial adhesion significantly (～81% at 15.0 μA cm^?2). The positive current did not significantly inhibit adhesion (<20% at 15.0 μA cm^?2), compared to the nonpolarized case. The alternating current had a similar effect as the negative current on preventing bacterial adhesion, but it also exhibited bactericidal effects, making it the most suitable method for bacterial adhesion control.     

Living in the fast lane: effects of cost of locomotion on foraging behaviour in juvenile Atlantic salmon

Stream-dwelling, juvenile Atlantic salmon, Salmo salar L., feed mainly on drifting invertebrates, usually by swimming upstream from a stationary position to intercept individual prey items. Laboratory experiments tested the prediction that individual salmon should reduce the distance over which they would travel (attack distance) to intercept drifting food items as the energy cost of swimming increases with increasing current velocity. Attack distance varied inversely with current velocity as expected. The fish's average speed of upstream movement relative to the substrate remained constant and the duration of individual attacks therefore declined as current velocity increased. Calculated reaction distances and a second ecperiment using tethered prey drifting at speeds independent of current velocity confirmed that these relationships were due to fish actually delaying attacks on perceived prey for longer periods as current velocity increased. Using estimated metabolic rates for burst swimming, it appears that energy expenditure per attack varies little with current velocity. Therefore, by reducing their reaction and attack distances in response to increasing current velocity, the fish reduced their energy cost of travel per attack.     

Flow-activated chloride channels in vascular endothelium. Shear stress sensitivity, desensitization dynamics, and physiological implications

Although activation of outward rectifying Cl(-) channels is one of the fastest responses of endothelial cells (ECs) to shear stress, little is known about these channels. In this study, we used whole-cell patch clamp recordings to characterize the flow-activated Cl(-) current in bovine aortic ECs (BAECs). Application of shear stress induced rapid development of a Cl(-) current that was effectively blocked by the Cl(-) channel antagonist 5-nitro-2-(3-phenopropylamino)benzoic acid (100 microM). The current initiated at a shear stress as low as 0.3 dyne/cm(2), attained its peak within minutes of flow onset, and saturated above 3.5 dynes/cm(2) approximately 2.5-3.5-fold increase over pre-flow levels). The Cl(-) current desensitized slowly in response to sustained flow, and step increases in shear stress elicited increased current only if the shear stress levels were below the 3.5 dynes/cm(2) saturation level. Oscillatory flow with a physiological oscillation frequency of 1 Hz, as occurs in disturbed flow zones prone to atherosclerosis, failed to elicit the Cl(-) current, whereas lower oscillation frequencies led to partial recovery of the current. Nonreversing pulsatile flow, generally considered protective of atherosclerosis, was as effective in eliciting the current as steady flow. Measurements using fluids of different viscosities indicated that the Cl(-) current is responsive to shear stress rather than shear rate. Blocking the flow-activated Cl(-) current abolished flow-induced Akt phosphorylation in BAECs, whereas blocking flow-sensitive K(+) currents had no effect, suggesting that flow-activated Cl(-) channels play an important role in regulating EC flow signaling.     

Effects of electrical currents on the absorption of water by eggs of Nereis limbata

Unfertilized eggs of the marine worm Nereis limbata subjected to electrical currents (direct or alternating) undergo remarkable changes. Certain minute granules just inside the surface of the egg absorb water and swell to more than 300 times their original size and thereby produce a mass of jelly which surrounds the egg with a zone about as wide as the original diameter of the egg. The amount of direct current is too small to produce any change of color in eggs stained with neutral red. In direct current the jelly appears first on the side toward the anode and moves toward the anode. In alternating current it appears on opposite sides facing the electrodes. It might be thought that the current changes the chemical character of the granules so that they are able to absorb very large quantities of water but this seems unlikely. If the current is shut off after 1 minute the swelling continues. This might be explained on the ground that each jelly precursor granule is covered with a waterproof film which is removed by the current. It does not seem probable that the effect is due to heat produced by the current since the exposure is so short. It seems possible that the current may strip off micelles from the waterproof covering of the granules and allow water to penetrate. The fact that alternating current is more effective than direct current might be explained on the ground that the egg may be represented as a capacity in parallel with a resistance so constituted that relatively little direct current can enter. The non-aqueous film which covers the surface of the protoplasm appears to be liquid rather than solid.     

Effects of manganese chloride on the outward currents in frog atrial trabeculae

The effects of MnCl2 on outward currents in frog atrial muscle were investigated under voltage-clamp conditions. MnCl2 (3 mmol/L), which completely abolished the slow inward current, produced a decrease in the outward background current (Ib) at potentials positive to -50 mV. The delayed outward current (Ix, time dependent) was not altered by Mn. "Isochronic activation curves" for Ix and decay of current tails at -40 mV remained unaffected after Mn. Effects on Ib probably reflect a decrease in IK1 related to the decrease in Ca influx as well as a reduction in the Na-Ca exchange current.     

Characterization of nifedipine-resistant calcium current in neonatal rat ventricular cardiomyocytes

Calcium current was recorded from ventricular cardiomyocytes of rats at various stages of postnatal development using the whole cell patch-clamp technique. In cultured 3-day-old neonatal cells, the current carried by Ca(2+) or Ba(2+) (5 mM) was not completely inhibited by 2 microM nifedipine. A residual current was activated in the same voltage range as the L-type, nifedipine-sensitive Ca(2+) current, but its steady-state inactivation was negatively shifted by 16 mV. This nifedipine-resistant calcium current was not further inhibited by other organic calcium current antagonists such as PN200-110, verapamil, and diltiazem nor by nickel, omega-conotoxin, or tetrodotoxin. It was completely blocked by cadmium and increased by isoproterenol and forskolin. This current was >20% of total calcium current in ventricular myocytes freshly isolated from neonatal rats, and it decreased during postnatal maturation, disappearing at the adult stage. This suggests that this current could be caused by an isoform of the L-type calcium channel expressed in a way that reflects the developmental stage of the rat heart.     

Is there an A-type K+ current in guinea pig ventricular myocytes?

A unique transient outward K(+) current (I(to)) has been described to result from the removal of extracellular Ca(2+) from ventricular myocytes of the guinea pig (15). This study addressed the question of whether this current represented K(+)-selective I(to) or the efflux of K(+) via L-type Ca(2+) channels. This outward current was inhibited by Cd(2+), Ni(2+), Co(2+), and La(3+) as well as by nifedipine. All of these compounds were equally effective inhibitors of the L-type Ca(2+) current. The current was not inhibited by 4-aminopyridine. Apparent inhibition of the outward current by extracellular Ca(2+) was shown to result from the displacement of the reversal potential of cation flux through L-type Ca(2+) channels. The current was found not to be K(+) selective but also permeant to Cs(+). The voltage dependence of inactivation of the outward current was identical to that of the L-type Ca(2+) current. It is concluded that extracellular Ca(2+) does not mask an A-type K(+) current in guinea pig ventricular myocytes.     

Facile technique of protein precipitation by application of electric current

Summary Precipitation of proteins has been achieved following passage of direct electric current in various protein solutions. Application of as low as 3 V of electric current showed precipitation but the rate increased with increase in electric current. With 9 V there was more than 85% precipitation of protein within 15 min. Precipitation occurred at a wide range of pH and temperature. Electrophoretic analysis of precipitated proteins show that they are not denatured by application of electric current. Proteins thus precipitated can be easily recovered by centrifugation.     

Endogenous electrical current leaves the limb and prelimb region of the Xenopus embryo

The electrical current pattern around the developing Xenopus laevis embryo was mapped with a vibrating probe. Current (taken as the movement of positive charge) was found to leave the emerging hind limb bud and to enter the gill region of the stage 47 embryo. The magnitude of the current leaving the limb was about 7 μA/cm² and the current entering the gill was about 60 μA/cm². Other regions of smaller outward current were found between the limb and gill. At stage 43, prior to the appearance of the limb bud, a highly localized region of outward current existed in the general area from which the bud would later emerge. The inward current was localized to the gill bud, as in the older embryo. The ionic basis of the currents could not be determined. In about one-third of the cases studied, the inward current was sodium sensitive since the removal of external sodium or the addition of amiloride reversably blocked the current. In the remaining cases, however, removal of sodium did not change (or else increased) the current. No other external ion (Ca²⁺, Mg²⁺, K⁺) could be identified as the current-carrying ion; the possibility of an outward movement of some anion such as Cl^? or HCO^?₃ remains.