Chara buckellii, a euryhaline charophyte from an unusual saline environment. II. Membrane potential and membrane conductance at steady state

1987 ◽  
Vol 65 (2) ◽  
pp. 222-229 ◽  
Author(s):  
Rosanne Hoffmann ◽  
Mary A. Bisson
Keyword(s):  
Steady State ◽  
Membrane Potential ◽  
Membrane Conductance ◽  
Proton Pumping ◽  
Membrane Properties ◽  
Saline Environment ◽  
High Ph ◽  
Threefold Increase ◽  
Cells Cultured ◽  
External K

Electrophysiological experiments were performed on internodal cells of Chara buckellii cultured in saline and freshwater media to see whether the membrane potential (Evo) and membrane conductance (Gm) are dominated by passive K+ conductance (K state), passive H+ conductance (H state), or active proton pumping (P state). Unlike other euryhaline charophytes, C. buckellii cells cultured under saline conditions were not dominated by any one state, showing little dependence on external K+ concentration and pH (pH°). Chara buckellii cells cultured in freshwater share some membrane properties with other freshwater charophytes. Freshwater cells appeared to be in the P state between pH° 5 and 7 as expected but never attained the H state usually observed at high pH° (> 10). Freshwater cells also showed a temporary, threefold increase in Gm at pH° 5, which could not be explained by an increase in passive Cl− or K+ conductance. Evidence consistent with an electrogenic Cl−/2H+ symport in freshwater-grown C. buckellii at pH° 5 and 7 is also presented.

scholarly journals The Influence of External Potassium on the Inactivation of Sodium Currents in the Giant Axon of the Squid, Loligo pealei

1969 ◽  
Vol 53 (6) ◽  
pp. 685-703 ◽  
Author(s):  
William J. Adelman ◽  
Yoram Palti
Keyword(s):  
Steady State ◽  
Membrane Potential ◽  
Membrane Conductance ◽  
Absolute Magnitude ◽  
Giant Axon ◽  
Membrane Potentials ◽  
Initial Transient ◽  
External Potassium ◽  
Loligo Pealei ◽  
External K

Isolated giant axons were voltage-clamped in seawater solutions having constant sodium concentrations of 230 mM and variable potassium concentrations of from zero to 210 mM. The inactivation of the initial transient membrane current normally carried by Na+ was studied by measuring the Hodgkin-Huxley h parameter as a function of time. It was found that h reaches a steady-state value within 30 msec in all solutions. The values of h∞, τh, αh,and ßh as functions of membrane potential were determined for various [Ko]. The steady-state values of the h parameter were found to be inversely related, while the time constant, τh, was directly related to external K+ concentration. While the absolute magnitude as well as the slopes of the h∞ vs. membrane potential curves were altered by varying external K+, only the magnitude and not the shape of the corresponding τh curves was altered. Values of the two rate constants, αh and ßh, were calculated from h∞ and τh values. αh is inversely related to [Ko] while ßh is directly related to [Ko] for hyperpolarizing membrane potentials and is independent of [Ko] for depolarizing membrane potentials. Hodgkin-Huxley equations relating αh and ßh to Em were rewritten so as to account for the observed effects of [Ko]. It is concluded that external potassium ions have an inactivating effect on the initial transient membrane conductance which cannot be explained solely on the basis of potassium membrane depolarization.

scholarly journals The influence of external cations and membrane potential on Ca-activated Na efflux in Myxicola giant axons.

1978 ◽  
Vol 71 (4) ◽  
pp. 453-466 ◽  
Author(s):  
R A Sjodin ◽  
R F Abercrombie
Keyword(s):  
Membrane Potential ◽  
Direct Action ◽  
Monovalent Cation ◽  
Coupling Ratio ◽  
Large Component ◽  
Experimental Conditions ◽  
Giant Axons ◽  
Threefold Increase ◽  
Na Efflux ◽  
External K

In microinjected Myxicola giant axons with elevated [Na]i, Na efflux was sensitive to Cao under some conditions. In Li seawater, sensitivity to Cao was high whereas in Na seawater, sensitivity to Cao was observed only upon elevation of [Ca]o above the normal value. In choline seawater, the sensitivity of Na efflux to Cao was less than that observed in Li seawater whereas Mg seawater failed to support any detectable Cao-sensitive Na efflux. Addition of Na to Li seawater was inhibitory to Cao-sensitive Na efflux, the extent of inhibition increasing with rising values of [Na]o. The presence of 20 mM K in Li seawater resulted in about a threefold increase in the Cao-activated Na efflux. Experiments in which the membrane potential, Vm, was varied or held constant when [K]o was changed showed that the augmentation of Ca-activated Na efflux by Ko was not due to changes in Vm but resulted from a direct action of K on activation by Ca. The same experimental conditions that favored a large component of Cao-activated Na efflux also caused a large increase in Ca influx. Measurements of Ca influx in the presence of 20 mM K and comparison with values of Ca-activated Na efflux suggest that the Na:Ca coupling ratio may be altered by increasing external [K]o. Overall, the results suggest that the Cao-activated Na efflux in Myxicola giant axons requires the presence of an external monovalent cation and that the order of effectiveness at a total monovalent cation concentration of 430 mM is K + Li greater than Li greater than Choline greater than Na.

scholarly journals Cat Heart Muscle in Vitro

1965 ◽  
Vol 48 (5) ◽  
pp. 933-948 ◽  
Author(s):  
Jon Goerke ◽  
Ernest Page
Keyword(s):  
Steady State ◽  
Membrane Potential ◽  
Papillary Muscle ◽  
Heart Muscle ◽  
Volume Ratio ◽  
K Concentration ◽  
Linear Manner ◽  
Cat Heart ◽  
External K

The exchange of cell K with K42, JK, has been measured in cat right ventricular papillary muscle under conditions of a steady state with respect to intracellular K concentration. Within the limits of the measurement, all of cell K exchanged at a single rate. Cells from small cats are smaller and have larger surface/volume ratios than cells from large cats. The larger surface/volume ratio results in larger flux values. JK increases in an approximately linear manner as the external K concentration is increased twentyfold, from 2.5 to 50 mM, at constant intracellular K concentration. The permeability for K ions, PK, calculated from the influx and membrane potential, remains very nearly constant over this range of external K concentrations. JK is not affected by replacement of O2 by N2, or by stimulated contractions at 60 per minute, but K influx decreases markedly in 10-5 M and 10-8 M ouabain.

Use of Membrane Potential Measurements to Study Mode of Action of Diclofop-Methyl

Weed Science ◽  
1994 ◽  
Vol 42 (2) ◽  
pp. 285-292 ◽  
Author(s):  
John P. Wright
Keyword(s):  
Membrane Potential ◽  
Energy Metabolism ◽  
Electrical Potential ◽  
Potential Gradient ◽  
Membrane Vesicles ◽  
Proton Pumping ◽  
Transport Processes ◽  
Membrane Properties ◽  
Electrical Potential Difference ◽  
Study Mode

All actively metabolizing cells have an electrical potential difference, negative on the interior, across their membranes. This electrochemical potential gradient is generated primarily by proton-pumping ATPases and provides the driving force for the transport of various ionic and neutral solutes. It is a key element in the energy metabolism of cells. Such factors as alteration of transport processes, energy metabolism, cytoplasmic pH, and membrane permeability have a direct effect on the magnitude of the membrane potential. In a brief survey, diclofop-methyl, diclofop, hydroxydiclofop, CGA 82725, haloxyfop-methyl, haloxyfop, bentazon, dinoseb, 4-hydroxy CIPC, and 2-hydroxy CIPC caused rapid depolarizations of the membrane potential of oat coleoptiles. Chlorsulfuron, dimethipin, propham, CIPC, dicamba, alachlor, metolachlor, napthalic anhydride, and paraquat had no measurable effects. The depolarizing effects of diclofop-reported earlier are used to illustrate the methods and interpretation of plant cell membrane potential measurements. Diclofop and diclofop-methyl affect the membrane properties of sensitive plant cells. Diclofop irreversibly depolarized the membrane potential and increased the proton permeability of sensitive cells but not resistant cells. It also increased the ATPase activity of isolated membrane vesicles. The mechanism through which diclofop exerted its effect is not fully understood. The equipment and techniques required for the intercellular recording of membrane potentials and resistance are described as well as the limitations of the techniques. A method not used in herbicide studies but with great potential for studies of herbicide interactions with membranes is patch clamp. A brief introduction to the methods will be given.

scholarly journals Sodium and potassium fluxes and membrane potential of human neutrophils: evidence for an electrogenic sodium pump.

1982 ◽  
Vol 79 (3) ◽  
pp. 453-479 ◽  
Author(s):  
L Simchowitz ◽  
I Spilberg ◽  
P De Weer
Keyword(s):  
Membrane Potential ◽  
Sodium Pump ◽  
Membrane Conductance ◽  
Human Neutrophils ◽  
Constant Field ◽  
Electrogenic Sodium Pump ◽  
Na Efflux ◽  
The Individual ◽  
Sodium And Potassium ◽  
External K

Sodium and potassium ion contents and fluxes of isolated resting human peripheral polymorphonuclear leukocytes were measured. In cells kept at 37 degrees C, [Na]i was 25 mM and [K]i was 120 mM; both ions were completely exchangeable with extracellular isotopes. One-way Na and K fluxes, measured with 22Na and 42K, were all approximately 0.9 meq/liter cell water . min. Ouabain had no effect on Na influx or K efflux, but inhibited 95 +/- 7% of Na efflux and 63% of K influx. Cells kept at 0 degree C gained sodium in exchange for potassium ([Na]i nearly tripled in 3 h); upon rewarming, ouabain-sensitive K influx into such cells was strongly enhanced. External K stimulated Na efflux (Km approximately 1.5 mM in 140-mM Na medium). The PNa/PK permeability ratio, estimated from ouabain insensitive fluxes, was 0.10. Valinomycin (1 microM) approximately doubled PK. Membrane potential (Vm) was estimated using the potentiometric indicator diS-C3(5); calibration was based on the assumption of constant-field behavior. External K, but not Cl, affected Vm. Ouabain caused a depolarization whose magnitude dependent on [Na]i. Sodium-depleted cells became hyperpolarized when exposed to the neutral exchange carrier monensin; this hyperpolarization was abolished by ouabain. We conclude that the sodium pump of human peripheral neutrophils is electrogenic, and that the size of the pump-induced hyperpolarization is consistent with the membrane conductance (3.7-4.0 microseconds/cm2) computed from the individual K and Na conductances.

Dependence of L-arginine accumulation on membrane potential in cultured human fibroblasts

1987 ◽  
Vol 253 (3) ◽  
pp. C391-C397 ◽  
Author(s):  
O. Bussolati ◽  
P. C. Laris ◽  
F. A. Nucci ◽  
V. Dall'Asta ◽  
N. Longo ◽  
...  
Keyword(s):  
Steady State ◽  
Membrane Potential ◽  
Transport System ◽  
Human Fibroblasts ◽  
Null Point ◽  
Facilitated Diffusion ◽  
Cultured Human Fibroblasts ◽  
The Relationship ◽  
Binding Component ◽  
External K

The cell-to-medium distribution ratios at steady state of L-arginine (RArg) and of the lipid-soluble cation tetraphenylphosphonium (RTPP) were studied as a function of the membrane potential (Em) in adult human fibroblasts. The relationship between RArg and Em was qualitatively similar to that of RTPP and Em. Quantitatively, RArg and RTPP differed in that 1) RTPP was much greater than RArg when Em was near zero, indicating a significant binding component in the uptake of TPP+ but not of L-arginine, and 2) after a correction for binding when Em is near zero, RTPP was still greater than RArg so that RT/F . ln RTPP exceeded RT/F . ln RArg by 10-25 mV. The pattern of the redistribution of accumulated TPP+ and arginine after an alteration of Em was identical. In null-point experiments, the external [K+] for which there were no changes in cellular TPP+ or L-arginine in the presence of high valinomycin (the null points) were very similar for the two probes. Em calculated from the null-point measurements (-70(-)-80 mV) was also very similar to RT/F . ln RArg and thus smaller than RT/F.ln RTPP. It was concluded that 1) there was an additional TPP+ binding as cellular [TPP] rose in response to more negative membrane potentials, 2) the transport system for L-arginine in these cells (system y+) operates as a facilitated diffusion system driven by the membrane potential, and 3) in some circumstances, L-arginine could be employed as a probe of Em.

Acidification of rat liver lysosomes: quantitation and comparison with endosomes

1993 ◽  
Vol 265 (4) ◽  
pp. C901-C917 ◽  
Author(s):  
R. W. Van Dyke
Keyword(s):  
Steady State ◽  
Membrane Potential ◽  
Rat Liver ◽  
Proton Pumps ◽  
Intact Cells ◽  
Early Endosomes ◽  
Medium Permeability ◽  
Electrogenic Proton Pump ◽  
Secondary Lysosomes ◽  
Permeability Studies

Both lysosomes and endosomes are acidified by an electrogenic proton pump, although studies in intact cells indicate that the steady-state internal pH (pHi) of lysosomes is more acid than that of endosomes. We undertook the present study to examine in detail the acidification mechanism of purified rat liver secondary lysosomes and to compare it with that of a population of early endosomes. Both endosomes and lysosomes exhibited ATP-dependent acidification, but proton influx rates were 2.4- to 2.7-fold greater for endosomes than for lysosomes because of differences in both buffering capacity and acidification rates, suggesting that endosomes exhibited greater numbers or rates of proton pumps. Lysosomes, however, exhibited a more acidic steady-state pHi due in part to a slower proton leak rate. Changes in medium Cl- increased acidification rates of endosomes more than lysosomes, and the lysosome ATP-dependent interior-positive membrane potential was only partially eliminated by high-Cl- medium. Permeability studies suggested that lysosomes were less permeable to Na+, Li+, and Cl- and more permeable to K+ and PO4(2-) than endosomes. Na-K-adenosine-triphosphatase did not appear to regulate acidification of either vesicle type. Endosome and lysosome acidification displayed similar inhibition profiles to N-ethylmaleimide, dicyclohexyl-carbodiimide, and vanadate, although lysosomes were somewhat more sensitive [concentration producing 50% maximal inhibition (IC50) 1 nM] to bafilomycin A1 than endosomes (IC50 7.6 nM). Oligomycin (1.5-3 microM) stimulated lysosome acidification due to shunting of membrane potential. Overall, acidification of endosomes and lysosomes was qualitatively similar but quantitatively somewhat different, possibly related to differences in the density or rate of proton pumps as well as vesicle permeability to protons, anions, and other cations.

Effects of ouabain on background and voltage-dependent currents in canine colonic myocytes

1990 ◽  
Vol 259 (3) ◽  
pp. C402-C408 ◽  
Author(s):  
E. P. Burke ◽  
K. M. Sanders
Keyword(s):  
Membrane Potential ◽  
Potential Gradient ◽  
Circular Muscle ◽  
Input Resistance ◽  
Pump Current ◽  
Muscle Layer ◽  
Membrane Properties ◽  
Resting Potential ◽  
Voltage Dependent ◽  
In Cells

Previous studies have suggested that the membrane potential gradient across the circular muscle layer of the canine proximal colon is due to a gradient in the contribution of the Na(+)-K(+)-ATPase. Cells at the submucosal border generate approximately 35 mV of pump potential, whereas at the myenteric border the pump contributes very little to resting potential. Results from experiments in intact muscles in which the pump is blocked are somewhat difficult to interpret because of possible effects of pump inhibitors on membrane conductances. Therefore, we studied isolated colonic myocytes to test the effects of ouabain on passive membrane properties and voltage-dependent currents. Ouabain (10(-5) M) depolarized cells and decreased input resistance from 0.487 +/- 0.060 to 0.292 +/- 0.040 G omega. The decrease in resistance was attributed to an increase in K+ conductance. Studies were also performed to measure the ouabain-dependent current. At 37 degrees C, in cells dialyzed with 19 mM intracellular Na+ concentration [( Na+]i), ouabain caused an inward current averaging 71.06 +/- 7.49 pA, which was attributed to blockade of pump current. At 24 degrees C or in cells dialyzed with low [Na+]i (11 mM), ouabain caused little change in holding current. With the input resistance of colonic cells, pump current appears capable of generating at least 35 mV. Thus an electrogenic Na+ pump could contribute significantly to membrane potential.

Electrogenic Na+ Transport in a Crustacean Coxal Receptor

1979 ◽  
Vol 78 (1) ◽  
pp. 29-45
Author(s):  
MAURIZIO MIROLLI
Keyword(s):  
Steady State ◽  
Membrane Potential ◽  
Input Resistance ◽  
Scylla Serrata ◽  
Partial Action ◽  
State Response ◽  
Electrogenic Na Pump ◽  
K Concentration ◽  
Low K ◽  
In Cells

1. The response of the coxal receptors of the crab Scylla serrata to step stretches consisted of a partial action potential, Vα, followed by a steady-state depolarization, V8. The input resistance of the fibre was reduced during V8. 2. In the absence of stimulation, the dendrites of the receptors depolarized when external Na+ was substituted with choline or Li+, and when the external K+ concentration was increased or decreased. The dendrites also depolarized when ouabain was added to the saline. 3. The amplitude of both Vα and V8 was dependent on external Na+. In cells which were depolarized by ouabain, the amplitude of V8 increased when the K+ concentration of the saline was reduced. 4. V8 was followed by a small, but long-lasting, after-potential which was depolarizing when the membrane potential was between −70 and −60 mV. In cells depolarized by ouabain or by low K+ saline, the after-potential became hyperpolarizing. 5. When trains of brief stretches (each 5 ms in duration) were used as stimuli, the cells responded with trains of Vα responses. During this tetanic stimulation the cells hyperpolarized; cessation of the stimulus train was followed by a long-lasting hyperpolarization (PTH). 6. PTH was abolished in Li+ saline, in low K+ saline, and in the presence of ouabain. In control or in low K+ saline, PTH was not accompanied by a decrease in the input resistance of the fibres. 7. It is concluded that an electrogenic Na+ pump (or equivalent process) contributes a substantial fraction of the membrane potential of the unstimulated coxal receptors. Pump activity could be increased by Na+-loading the distal part of the cells with trains of Vα responses. By contrast, during the steady-state response to stretch, the pump was not activated.

Voltage behavior along the irregular dendritic structure of morphologically and physiologically characterized vagal motoneurons in the guinea pig

1990 ◽  
Vol 63 (2) ◽  
pp. 333-346 ◽  
Author(s):  
R. Nitzan ◽  
I. Segev ◽  
Y. Yarom
Keyword(s):  
Steady State ◽  
Guinea Pig ◽  
Time Constant ◽  
Dendritic Structure ◽  
Input Resistance ◽  
Membrane Properties ◽  
Small Cells ◽  
Physiological Data ◽  
Motor Nucleus ◽  
Cable Length

1. Intracellular recordings from neurons in the dorsal motor nucleus of the vagus (vagal motoneurons, VMs) obtained in the guinea pig brain stem slice preparation were used for both horseradish peroxidase (HRP) labeling of the neurons and for measurements of their input resistance (RN) and time constant (tau 0). Based on the physiological data and on the morphological reconstruction of the labeled cells, detailed steady-state and compartmental models of VM were built and utilized to estimate the range of membrane resistivity, membrane capacitance, and cytoplasm resistivity values (Rm, Cm, and Ri, respectively) and to explore the integrative properties of these cells. 2. VMs are relatively small cells with a simple dendritic structure. Each cell has an average of 5.3 smooth (nonspiny), short (251 microns) dendrites with a low order (2) of branching. The average soma-dendritic surface area of VMs is 9,876 microns 2. 3. Electrically, VMs show remarkably linear membrane properties in the hyperpolarizing direction; they have an average RN of 67 +/- 23 (SD) M omega and a tau 0 of 9.4 +/- 4.1 ms. Several unfavorable experimental conditions precluded the possibility of faithfully recovering ("peeling") the first equalizing time constant (tau 1) and, thereby, of estimating the electrotonic length (Lpeel) of VMs. 4. Reconciling VM morphology with the measured RN and tau 0 through the models, assuming an Ri of 70 omega.cm and a spatially uniform Rm, yielded an Rm estimate of 5,250 omega.cm2 and a Cm of 1.8 microF/cm2. Peeling theoretical transients produced by these models result in an Lpeel of 1.35. Because of marked differences in the length of dendrites within a single cell, this value is larger than the maximal cable length of the dendrites and is twice as long as their average cable length. 5. The morphological and physiological data could be matched indistinguishably well if a possible soma shunt (i.e., Rm, soma less than Rm, dend) was included in the model. Although there is no unique solution for the exact model Rm, a general conclusion regarding the integrative capabilities of VM could be drawn. As long as the model is consistent with the experimental data, the average input resistance at the dendritic terminals (RT) and the steady-state central (AFT----S) and peripheral (AFS----T) attenuation factors are essentially the same in the different models. With Ri = 70 omega.cm, we calculated RT, AFS----T, and AFT----S to be, on the average, 580 M omega, 1.1, and 13, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)

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