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H Lecar

Publications and source records attributed to H Lecar.

36 records · Page 2Linked to original sources

Acetylcholine-induced current fluctuations in tissue-cultured muscle cells under voltage clamp.

Acetylcholine applied ionophoretically to chick skeletal muscle cells grown in tissue culture produces membrane current fluctuations. Cells treated with vinblastine are transformed to a roughly spherical shape. Such transformed cells can be voltage-clamped with microelectrodes. The frequency spectrum of the current fluctuations at fixed voltage obeys a relation of the Lorentz form. From analysis of the current noise, the conductance of a single ionic channel is estimated to be 39 pmho at a temperature of 28 degrees C, and increases with increasing temperature, exhibiting a Q10 of 1.7. The relaxation time for the channel conductance is more sharply temperature dependent, showing a Q10 of approximately 5. These results are in agreement with the picture of acetylcholine-activated ionic channels determined from experiments on frog end plate (Anderson and Stevens, 1973). The relaxation time for carbachol activation is shorter than for acetylcholine, and appears to be more temperature sensitive.

Acetylcholine↗

Determination of the resistance in series with the membranes of giant axons.

Measurements of the resistance in series with the excitable membrane for giant axons of two different phylla (the squid Loligo pealii and the marine worm Myxicola infundibulum) were obtained. Efforts were made to take into account the errors introduced by the finite rise-time of the measuring apparatus. The series resistance value, obtained very quickly by the method described, may be used in setting the compensation potentiometer to offset this resistance in voltage-clamp measurements. Estimates of the resistance of the periaxonal tissue layer were made. Analyses were done on some of the problems involved in attempting to make an unambiguous determination of the series resistance.

Animals↗

Squid axon membrane response to white noise stimulation.

The current from a white noise generator was applied as a stimulus to a space-clamped squid axon in double sucrose gap. The membrane current and the voltage response of the membrane were then amplified, recorded on magnetic tape, and the stimulus was cross-correlated with the response. With subthreshold stimuli, a cross-correlation function resembling that obtained from a resonant parallel circuit is obtained. As the intensity of the input noise is increased, the cross-correlation function resembles that obtained from a less damped oscillatory circuit. When the noise intensity is further increased so that an appreciable frequency of action potentials is observed, an additional component appears in the experimental cross-correlogram. The subthreshold cross-correlogram is analyzed theoretically in terms of the linearized Hodgkin-Huxley equations. The subthreshold axon approximates a parallel resonant circuit. The circuit parameters are temperature dependent, with resonant frequency varying from approximately 100 Hz at 10 degrees C to approximately 250 Hz at 20 degrees C. The Q(10) of the resonant frequency is equal to 1.9. These values are in agreement with values found previously for subthreshold oscillations following a single action potential.

Acoustic Stimulation↗

Kinetics of the opening and closing of individual excitability-inducing material channels in a lipid bilayer.

The kinetics of the opening and closing of individual ion-conducting channels in lipid bilayers doped with small amounts of excitability-inducing material (EIM) are determined from discrete fluctuations in ionic current. The kinetics for the approach to steady-state conductance during voltage clamp are determined for lipid bilayers containing many EIM channels. The two sets of measurements are found to be consistent, verifying that the voltage-dependent conductance of the many-channel EIM system arises from the opening and closing of individual EIM channels. The opening and closing of the channels are Poisson processes. Transition rates for these processes vary exponentially with applied potential, implying that the energy difference between the open and closed states of an EIM channel is linearly proportional to the transmembrane electric field. A model incorporating the above properties of the EIM channels predicts the observed voltage dependence of ionic conductance and conductance relaxation time, which are also characteristic of natural electrically excitable membranes.

Cholesterol↗

Ion transport through excitability-inducing material (EIM) channels in lipid bilayer membranes.

Two different methods were used to determine the relative permeability and the voltage-dependent conductance of several different cations in excitability-inducing material (EIM)-doped lipid bilayers. In one method, the conductances of individual channels were measured for Li, Na, K, Cs, NH(4), and Ca, and in the other method biionic potentials of a membrane with many channels were measured for Li, Na, K, Cs, and Rb. The experimental results for the two methods are in agreement. The relative permeabilities are proportional to the ionic mobilities in free aqueous solution. The voltage dependence of the conductance is the same for all cations measured.

Cesium↗

Detection of molecular motion in lyophilized myelin by nuclear magnetic resonance.

Proton resonance spectroscopy was used to determine the state of the hydrocarbon regions in lyophilized and resuspended samples of nerve myelin. Measurements of the resonance line width indicate considerable freedom of motion within the hydrocarbon moiety of the myelin samples. Sharp thermal transitions of the line width were observed, suggesting that lyophilized myelin is in a liquid crystalline state.

Animals↗

Theory of threshold fluctuations in nerves. I. Relationships between electrical noise and fluctuations in axon firing.

Relations describing threshold fluctuation phenomena in nerves are derived by calculating the approximate response of the Hodgkin-Huxley (HH) axon to electrical noise. We use FitzHugh's reduced phase space approximation and describe the dynamics of a noisy nerve by a two-dimensional brownian motion. The theory predicts the functional form and parametric dependence of the relation between probability of firing and stimulus strength. Expressions are also obtained for the firing probability as a function of stimulus duration and for the distribution of latency times as a function of stimulus strength.

Action Potentials↗

Theory of threshold fluctuations in nerves. II. Analysis of various sources of membrane noise.

Threshold fluctuations in axon firing can arise as a result of electrical noise in the excitable membrane. A general theoretical expression for the fluctuations is applied to the analysis of three sources of membrane noise: Johnson noise, excess 1/f noise, and sodium conductance fluctuations. Analytical expressions for the width of the firing probability curve are derived for each of these noise sources. Specific calculations are performed for the node of Ranvier of the frog, and attention is given to the manner in which threshold fluctuations are affected by variations of temperature, ion concentrations, and the application of various drugs. Comparison with existing data suggests that threshold fluctuations can best be explained by sodium conductance fluctuations. Additional experiments directed at distinguishing among the various noise sources are proposed.

Action Potentials↗

The nature of the negative resistance in bimolecular lipid membranes containing excitability-inducing material.

When sufficiently small amounts of excitability-inducing material (EIM) are added to a bimolecular lipid membrane, the conductance is limited to a few discrete levels and changes abruptly from one level to another. From our study of these fluctuations, we have concluded that the EIM-doped bilayer contains ion-conducting channels capable of undergoing transitions between two states of different conductance. The difference in current between the "open" and "closed" states is directly proportional to the applied membrane potential, and corresponds to a conductance of about 3 x 10(-10) ohm(-1). The fraction of the total number of channels that is open varies from unity to zero as a function of potential. The voltage-dependent opening and closing of channels explains the negative resistance observed for bimolecular lipid membranes treated with greater amounts of EIM.

Electrophysiology↗

Ammonium ion currents in the squid giant axon.

Voltage-clamp studies on intact and internally perfused squid giant axons demonstrate that ammonium can substitute partially for either sodium or potassium. Ammonium carries the early transient current with 0.3 times the permeability of sodium and it carries the delayed current with 0.3 times the potassium permeability. The conductance changes observed in voltage clamp show approximately the same time course in ammonium solutions as in the normal physiological solutions. These ammonium ion permeabilities account for the known effects of ammonium on nerve excitability. Experiments with the drugs tetrodotoxin (TTX) and tetraethyl ammonium chloride (TEA) demonstrate that these molecules block the early and late components of the current selectively, even when both components are carried by the same ion, ammonium.

Action Potentials↗

Removal of potassium negative resistance in perfused squid giant axons.

Squid giant axons, internally and externally perfused with solutions having potassium as the only cation, exhibit an approximately linear steady-state current-voltage relation. When small amounts of calcium and magnesium are present in the external potassium solution, the current-voltage curve is markedly nonlinear, exhibiting the rectification and negative resistance which have been observed for intact axons in isosmotic potassium solutions. The effects of perfusion and removal of external divalent cations are interpreted in terms of two components of current, a linear component and a nonlinear time-varying component. The former is increased and the latter diminished by the removal of the external divalent cations.

Animals↗

Single postsynaptic channel currents in tissue cultured muscle.

Some of the most compelling evidence for the existence of ionic channels in cell membranes comes from direct recording of quantised current jumps generated by the opening and closing of individual channels. Single-channel jumps have been extensively studied for lipid bilayer membranes doped with various channel-forming additives. Recently agonist-induced single-channel currents were detected in denervated frog muscle by use of extracellular electrodes, which can isolate the current from a small area of membrane. The current jumps provide a means for the direct test of many of the inferences about ionic channels which have come from electrical noise analysis. In this report we present measurements of single-channel currents induced by the agonist carbamylcholine in tissue-cultured mammalian muscle. These measurements confirm the earlier noise studies on tissue culture preparations. Recordings of single-channel currents induced by the agonist, suberyldicholine, in avian muscle are presented by Nelson and Sachs.

Animals↗