Discontinuous feedback amplifier: principles and application to the measurement of transmembrane potentials and currents of frog atrial fibers

The study and achievement of a discontinuous feedback amplifier to measure membrane potentials and currents in frog atrial fibres using the double sucrose gap technique was achieved. It was shown that, with the present device, the effects of the resistance in series with the membrane resistance and the membrane capacity on the measure of cardiac membrane potentials and fast currents are markedly reduced.     

Voltage Clamp of Cardiac Muscle in a Double Sucrose Gap: A Feasibility Study

A method of stabilizing the membrane potential of a small area of cardiac muscle membrane and the limitations of this method are described. Tiny bundles or strands, approximately 80 μm in diameter, of electrically interconnected fibers from the ventricles of rabbit hearts were used in a double sucrose gap. Current records associated with step changes in voltage were complicated by two capacitive surges of current of nodal and nonnodal origin and large “leakage” currents of nonnodal origin resulting mainly from the multifibered nature of the preparation and emphasized by the method. The transient, inward membrane currents in response to moderate depolarizing steps in command potential had the same duration as the upstroke of the action potential. In good runs, currents were smooth and free from notches. These initial currents behaved qualitatively like the initial sodium currents in squid axon and in other excitable membranes. A fraction of the initial sodium current persisted at least as long as 300 ms. The relationship between peak initial current and voltage was graded and linear in the positive direction. In the negative region the relationship was often very steep, indicating insufficient voltage control of all the membranes despite the squareness of the voltage record. Other indications of inadequacy of control could occur and thus even with this optimum preparation of cardiac muscle it was not feasible to analyze quantitatively either the initial or the prolonged sodium currents.     

Voltage-dependent membrane ionic currents in lamprey striated muscle

Membrane ionic currents in striated muscle bundles of lamprey suction apparatus were recorded using a double sucrose gap technique. Transmembrane currents in a single muscle fiber and a fiber bundle in the frog were compared so as to check the validity of current measurement in multicell preparations. It was found that fast inward sodium currents arise in the lamprey muscle membrane in response to depolarization together with a delayed outward potassium current, with steady-state characteristics resembling those of membrane currents in frog muscle. The only difference consisted of a flatter curve for steady-state inactivation of potassium current, probably indicative of greater density of potassium channels. Both the changes in reversal potential and the speed of potassium current deactivation occurring during protracted stimuli point to the presence of two fractions in this current. No functioning voltage-dependent calcium channels are found in the lamprey muscle membrane.I. M. Sechenov Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 18, No. 5, pp. 629–636, September–October, 1986.     

The Potential Distribution and the Short-Circuiting Factor in the Sucrose Gap

The sucrose gap technique, though widely employed in many tissues, could not be used for quantitative measurements of the membrane potential, because the value of the short-circuiting factor and the influence of junction potential on the recorded potential difference were unknown. The formula that relates the recorded potential to the true resting membrane potential was found by application of the cable equations to a core conductor placed in a system with three different media, e.g. Ringer, sucrose, and KCl. The formula shows that the potential difference recorded over the sucrose insulator depends on the extracellular and the intracellular longitudinal resistances, the membrane resistance and the membrane potentials in each region, and on the junction potentials between the different media. The true membrane potential in the Ringer region can be calculated from the potential difference recorded after complete depolarization by KCl on one side of the preparation, if the longitudinal resistances, the membrane resistances, the extracellular potential in the sucrose, and the junction potential between Ringer and sucrose are determined by separate measurements.     

An Assessment of the Double Sucrose-Gap Voltage Clamp Technique as Applied to Frog Atrial Muscle

The homogeneity of voltage clamp control in small bundles of frog atrial tissue under double sucrose-gap voltage clamp conditions was assessed by intracellular microelectrode potential measurements from cells in the test node region. The microelectrode potential measurements demonstrated that (1) good voltage control of the impaled cell existed in the absence of the excitatory inward currents (e.g., during small depolarizing clamp pulses of 10-15 mV), (2) voltage control of the impaled cell was lost during either the fast or slow excitatory inward currents, and (3) voltage control of the impaled cell was regained following the inward excitatory currents. Under nonvoltage clamp conditions the transgap recorded action potential had a magnitude and waveform similar to the intracellular microelectrode recorded action potentials from cells in the test node. Transgap impedance measured with a sine-wave voltage of 1,000 Hz was about 63% of that measured either by a sine-wave voltage of 10 Hz or by an action potential method used to determine the longitudinal resistance through the sucrose-gap region. The action potential data in conjunction with the impedance data indicate that the extracellular resistance (R_s) through the sucrose gap is very large with respect to the longitudinal intracellular resistance (R_i); the frequency dependence of the transgap impedance suggests that at least part of the intracellular resistance is paralleled by a capacitance. The severe loss of spatial voltage control during the excitatory inward current raises serious doubts concerning the use of the double sucrose-gap technique to voltage clamp frog atrial muscle.     

Computing the Local Field Potential (LFP) from Integrate-and-Fire Network Models

Leaky integrate-and-fire (LIF) network models are commonly used to study how the spiking dynamics of neural networks changes with stimuli, tasks or dynamic network states. However, neurophysiological studies in vivo often rather measure the mass activity of neuronal microcircuits with the local field potential (LFP). Given that LFPs are generated by spatially separated currents across the neuronal membrane, they cannot be computed directly from quantities defined in models of point-like LIF neurons. Here, we explore the best approximation for predicting the LFP based on standard output from point-neuron LIF networks. To search for this best “LFP proxy”, we compared LFP predictions from candidate proxies based on LIF network output (e.g, firing rates, membrane potentials, synaptic currents) with “ground-truth” LFP obtained when the LIF network synaptic input currents were injected into an analogous three-dimensional (3D) network model of multi-compartmental neurons with realistic morphology, spatial distributions of somata and synapses. We found that a specific fixed linear combination of the LIF synaptic currents provided an accurate LFP proxy, accounting for most of the variance of the LFP time course observed in the 3D network for all recording locations. This proxy performed well over a broad set of conditions, including substantial variations of the neuronal morphologies. Our results provide a simple formula for estimating the time course of the LFP from LIF network simulations in cases where a single pyramidal population dominates the LFP generation, and thereby facilitate quantitative comparison between computational models and experimental LFP recordings in vivo.     

Carrier Model for Active Transport of Ions across a Mosaic Membrane

The central purpose of this paper is to elucidate in a well defined system the meaning of certain phenomena and concepts associated with the active transport of ions. To this end a specific model for a carrier system which actively transports a single ionic species is analyzed and discussed in detail. It is assumed in this model that the carrier-mediated ionic transport occurs in regions of the membrane physically separate from those regions in which free ionic movement takes place,—coupling between the active and passive regions of the membrane occurring through local current flows. The model is seen to display the following characteristics: (a) Starting from identical solutions on the two sides of the membrane, there is produced a redistribution of ions; (b) with identical solutions on the two sides of the membrane there exists a potential difference across the membrane, i.e., the “pump” is electrogenic; (c) the “short circuit” current for symmetrical solutions is equal to the flux of the neutral ion carrier complex; (d) the rate of active transport (and hence of metabolism) is dependent on the ionic concentrations in the surrounding solutions. Throughout the paper comparison is made between features of the model and properties displayed by biological active transport systems, but there is no claim of an identity between the two.     

Potassium ion accumulation in a periaxonal space and its effect on the measurement of membrane potassium ion conductance

Summary Potassium currents of various durations were obtained from squid giant axons voltage-clamped in artificial seawater solutions containing sufficient tetrodotoxin to block the sodium conductance completely. From instantaneous potassium current-voltage relations, the reversal potentials immediately at the end of these currents were determined. On the basis of these reversal potential measurements, the potassium ion concentration gradient across the membrane was shown to decrease as the potassium current duration increased. The kinetics of this change was shown to vary monotonically with the potassium ion efflux across the membrane estimated from the integral over time of the potassium current divided by the Faraday, and to be independent of both the external sodium ion concentration and the presence or absence of membrane series resistance compensation. It was assumed that during outward potassium current flow, potassium ions accumulated in a periaxonal space bounded by the membrane and an external diffusion barrier. A model system was used to describe this accumulation as a continuous function of the membrane currents. On this basis, the mean periaxonal space thickness and the permeability of the external barrier to K⁺ were found to be 357 Å and 3.21×10^–4 cm/sec, respectively. In hyperosmotic seawater, the value of the space thickness increased significantly even though the potassium currents were not changed significantly. Values of the resistance in series with the membrane were calculated from the values of the permeability of the external barrier and these values were shown to be roughly equivalent to series resistance values determined by current clamp measurements. Membrane potassium ion conductances were determined as a function of time and voltage. When these were determined from data corrected for the potassium current reversal potential changes, larger maximal potassium conductances were obtained than were obtained using a constant reversal potential. In addition, the potassium conductance turn-on with time at a variety of membrane potentials was shown to be slower when potassium conductance values were obtained using a variable reversal potential than when using a constant reversal potential.     

Reexamination of the double sucrose gap technique for the study of lobster giant axons. Theory and experiments

The double sucrose gap technique for the study of lobster giant axons has been reexamined. The leakage behavior of the system cannot be successfully modeled by conventional sucrose gap theory, but is accounted for by the McGuigan-Tsien model that takes into account the cable properties of membrane under sucrose. The facts of high-leakage conductance and the ability to maintain large resting potentials in the face of low sucrose gap resistance lead to a hypothesis that membrane resistance under sucrose is very low because of a large negative surface potential. Computer simulations of the leakage behavior of the conventional gap model and the McGuigan-Tsien model were compared with experimental measurements on lobster axons using normal sucrose or sucrose doped with Na+, Ca2+ or La3+ ions. As the concentration of doping ion increased, the leakage rose, but the species of doping ion had more influence on leakage than gap resistance. At equal gap resistance, leakage decreased with an increase in valence of the doping species. Leakage was even lower in La-doped sucrose at 20 M omega gap resistance than in normal sucrose at 200 M omega gap resistance. Resting potentials decreased with decreasing gap resistance and increasing valence of the doping species. Resting potential behavior was successfully simulated with a hybrid model consisting of a point node flanked by infinite cables and a shunt between ground and the voltage-measuring pool. The data support the hypothesis that the membrane resistance under sucrose is low and that it can be raised by doping the sucrose with multivalent cations, with La3+ being particularly effective.(ABSTRACT TRUNCATED AT 250 WORDS)     

Quantifying Auditory Temporal Stability in a Large Database of Recorded Music

“Moving to the beat” is both one of the most basic and one of the most profound means by which humans (and a few other species) interact with music. Computer algorithms that detect the precise temporal location of beats (i.e., pulses of musical “energy”) in recorded music have important practical applications, such as the creation of playlists with a particular tempo for rehabilitation (e.g., rhythmic gait training), exercise (e.g., jogging), or entertainment (e.g., continuous dance mixes). Although several such algorithms return simple point estimates of an audio file’s temporal structure (e.g., “average tempo”, “time signature”), none has sought to quantify the temporal stability of a series of detected beats. Such a method-a “Balanced Evaluation of Auditory Temporal Stability” (BEATS)–is proposed here, and is illustrated using the Million Song Dataset (a collection of audio features and music metadata for nearly one million audio files). A publically accessible web interface is also presented, which combines the thresholdable statistics of BEATS with queryable metadata terms, fostering potential avenues of research and facilitating the creation of highly personalized music playlists for clinical or recreational applications.     

Computer simulation of the effect of the nodal gap resistance on ionic current measurements in the Ranvier node membrane.

Results are presented of a computer simulation of the effect of the irreducible resistance introduced by the nodal gap, in series with the impedance of the axon membrane. A clamp potential is applied to a structure modeled as an electric circuit composed of a resistance in series with the membrane impedance, and modified nerve equations describing membrane currents are solved to predict the effect of nodal series resistance on these currents. These studies reveal changes in the absolute values and kinetics of the ionic currents (errors greater than 10-20%) for selected values of series resistance.     

The correction factors for sucrose gap measurements and their practical applications.

The distribution of extracellular and intracellular potential in the sucrose gap apparatus, previously established for a single fiber using the cable equations for a core conductor model (Jirounek and Straub, Biophys. J., 11:1, 1971), is obtained for a multifiber preparation. The exact equation is derived relating the true membrane potential change to the measured potential differences across the sucrose gap, the junction potentials between sucrose and physiological solution, the membrane potential in the sucrose region, and the electrical parameters of the preparation in each region of the sucrose gap. The extracellular potential distribution has been measured using a modified sucrose gap apparatus for the frog sciatic nerve and the rabbit vagus nerve. The results indicate a hyperpolarization of the preparations in the sucrose region, of 60--75 mV. The hyperpolarization is independent of the presence of junction potentials. The calculation of the correction terms in the equation relating the actual to the measured potential change is illustrated for the case of complete depolarization by KC1 on one side of the sucrose gap. The correction terms in the equation are given for various experimental conditions, and a number of nomographic charts are presented, by means of which the correction factors can be rapidly evaluated.     

Behavioral Characterization of the Hyperphagia Synphilin-1 Overexpressing Mice

Synphilin-1 is a cytoplasmic protein that has been shown to be involved in the control of energy balance. Previously, we reported on the generation of a human synphilin-1 transgenic mouse model (SP1), in which overexpression of human synphilin-1 resulted in hyperphagia and obesity. Here, behavioral measures in SP1 mice were compared with those of their age-matched controls (NTg) at two time points: when there was not yet a group body weight difference (“pre-obese”) and when SP1 mice were heavier (“obese”). At both time points, meal pattern analyses revealed that SP1 mice displayed higher daily chow intake than non-transgenic control mice. Furthermore, there was an increase in meal size in SP1 mice compared with NTg control mice at the obese stage. In contrast, there was no meal number change between SP1 and NTg control mice. In a brief-access taste procedure, both “pre-obese” and “obese“ SP1 mice displayed concentration-dependent licking across a sucrose concentration range similar to their NTg controls. However, at the pre-obese stage, SP1 mice initiated significantly more trials to sucrose across the testing sessions and licked more vigorously at the highest concentration presented, than the NTg counterparts. These group differences in responsiveness to sucrose were no longer apparent in obese SP1 mice. These results suggest that at the pre-obese stage, the increased trials to sucrose in the SP1 mice reflects increased appetitive behavior to sucrose that may be indicative of the behavioral changes that may contribute to hyperphagia and development of obesity in SP1 mice. These studies provide new insight into synphilin-1 contributions to energy homeostasis.     

Properties of internally perfused, voltage-clamped, isolated nerve cell bodies

The membrane properties of isolated neurons from Helix aspersa were examined by using a new suction pipette method. The method combines internal perfusion with voltage clamp of nerve cell bodies separated from their axons. Pretreatment with enzymes such as trypsin that alter membrane function is not required. A platinized platinum wire which ruptures the soma membrane allows low resistance access directly to the cell's interior improving the time resolution under voltage clamp by two orders of magnitude. The shunt resistance of the suction pipette was 10-50 times the neuronal membrane resistance, and the series resistance of the system, which was largely due to the tip diameter, was about 10(5) omega. However, the peak clamp currents were only about 20 nA for a 60-mV voltage step so that measurements of membrane voltage were accurate to within at least 3%. Spatial control of voltage was achieved only after somal separation, and nerve cell bodies isolated in this way do not generate all-or-none action potentials. Measurements of membrane potential, membrane resistance, and membrane time constant are equivalent to those obtained using intracellular micropipettes, the customary method. With the axon attached, comparable all-or-none action potentials were also measured by either method. Complete exchange of Cs+ for K+ was accomplished by internal perfusion and allowed K+ currents to be blocked. Na+ currents could then be blocked by TTX or suppressed by Tris-substituted snail Ringer solution. Ca2+ currents could be blocked using Ni2+ and other divalent cations as well as organic Ca2+ blockers. The most favorable intracellular anion was aspartate-, and the sequence of favorability was inverted from that found in squid axon.     

Electrical Characteristics of Tunicate Heart Cell Membranes and Nexuses

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 cm² and the membrane capacitance was about 1.6 µF per cm². 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 cm². It is concluded that the nexuses provide a low resistance pathway between cells and a transepithelial barrier.     

Whole-Cell K+ Currents across the Plasma Membrane of Tobacco Protoplasts from Cell-Suspension Cultures

The whole-cell configuration of the patch clamp technique was used to study both outward and inward ion currents across the plasma membrane of tobacco (Nicotiana tabacum) protoplasts from cell-suspension cultures. The ion currents across the plasma membrane were analyzed by the application of stepwise potential changes from a holding potential or voltage ramps. In all protoplasts, a voltage- and time-dependent outward rectifying current was present. The conductance increased upon depolarization of the membrane potential (to >0 mV) with a sigmoidal time course. The reversal potential of the outward current shifted in the direction of the K+ equilibrium potential upon changing the external K+ concentration. The outward current did not show inactivation. In addition to the outward rectifying current, in about 30% of the protoplasts, a time- and voltage-dependent inward rectifying current was present as well. The inward rectifying current activated upon hyperpolarization of the membrane potential (<-100 mV) with an exponential time course. The reversal potential of the inward conductance under different ionic conditions was close to the K+ equilibrium potential.     

When Feelings Arise with Meanings: How Emotion and Meaning of a Native Language Affect Second Language Processing in Adult Learners

To determine when and how L2 learners start to process L2 words affectively and semantically, we conducted a longitudinal study on their interaction in adult L2 learners. In four test sessions, spanning half a year of L2 learning, we monitored behavioral and ERP learning-related changes for one and the same set of words by means of a primed lexical-decision paradigm with L1 primes and L2 targets. Sensitivity rates, accuracy rates, RTs, and N400 amplitude to L2 words and pseudowords improved significantly across sessions. A semantic priming effect (e.g, prime “driver”facilitating response to target “street”) was found in accuracy rates and RTs when collapsing Sessions 1 to 4, while this effect modulated ERP amplitudes within the first 300 ms of L2 target processing. An overall affective priming effect (e.g., “sweet” facilitating”taste”) was also found in RTs and ERPs (posterior P1). Importantly, the ERPs showed an L2 valence effect across sessions (e.g., positive words were easier to process than neutral words), indicating that L2 learners were sensitive to L2 affective meaning. Semantic and affective priming interacted in the N400 time-window only in Session 4, implying that they affected meaning integration during L2 immersion together. The results suggest that L1 and L2 are initially processed semantically and affectively via relatively separate channels that are more and more linked contingent on L2 exposure.     

Origin of the long positive potential in the cat superior cervical ganglion

The sucrose gap technique was used to study the long positive potential (P-potential) in a curarized cat superior cervical ganglion. The frequency of stimulating the preganglionic trunk optimal for P-potential production was 30–40 impulses/sec at a stimulus series duration of 1 sec. Proserine in low concentrations (1–5 µg/ml) increased amplitude and especially duration of the P-potential. Atropine (0.5–2 µg/ml) blocked it completely. Adrenaline and noradrenaline (10–50 µg/ml inhibited both the negative potential (corresponding to the fast EPSP of neurons) and the P-potential in equal measure. The nature of dependence of P-potential amplitude on value of the membrane potential was also studied. It was found that the P-potential is inhibited in solutions with low potassium ion content, and that amplitude of the P-potential rises with an increase of intracellular sodium concentration. The rate of its increase rises with an increase of temperature. Under the influence of strophathin, the P-potential is inhibited. The data obtained support the hypothesis that the P-potential is determined by synaptic activation of the electrogenic sodium pump.A. A. Bogomol'ets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 3, No. 1, pp. 76–83, January–February, 1971.     

Oscillations in the electrical resistance of Nitellopsis obtusa nodes

Oscillatory changes of the electrical resistance across the nodal complex of Nitellopsis obtusa (Desv. in Lois) J. Gr. were observed in experiments performed for 40–150 min with the use of external electrodes and microelectrodes. Three main patterns of node resistance oscillations were similar to those found for membrane potential and resistance. The presented findings indicate an oscillatory behaviour of the plasmodesmata system at the node, which may be connected with e.g. pulsatile variations in the number of open plasmodesmata.