Electrical Evidence from Perfused and Intact Cells for Voltage-dependent K+ Channels in the Plasmalemma of Chara australis

1984 ◽  
Vol 11 (4) ◽  
pp. 303 ◽  
Author(s):  
P.T Smith
Keyword(s):  
Steady State ◽  
Negative Slope ◽  
Equilibrium Potential ◽  
Intact Cells ◽  
K Channels ◽  
Current Voltage ◽  
Voltage Dependent ◽  
Chara Australis ◽  
Opening And Closing ◽  
External K

The K+ conductance in Chara australis was studied using current-voltage relations (I- V curves) of intact cells and perfused plasmalemmas in conditions which minimized H+ conductance. The rapid I-V curves of perfused cells were dominated by K+ conductance, intersecting at the calculated K' equilibrium potential. The steady state I-V curves were non-linear; the point of greatest change in slope conductance has been called the 'switch potential'. At voltages more positive than the switch potential, K+ conductance increased with potential. At more negative voltages, K+ conductance was constant and low. In high external K+ concentrations the steady state I-V curves developed negative slope conductance near the switch potential. The switch potential behaved as a function of the KC equilibrium potential. During prolonged voltage pulses in perfused cells, the clamp current changed exponentially with time. The values o f t , were affected by the size of the voltage pulse and the external KT concentration. The results can be explained by time- and voltage-dependent K+ channels. It is suggested that the voltage sensor, which supposedly regulates the opening and closing of the Kt channels, measures a function of the membrane potential and the K+ equilibrium potential.

scholarly journals Steady-state current-voltage relationship of the Na/K pump in guinea pig ventricular myocytes.

1989 ◽  
Vol 94 (3) ◽  
pp. 511-537 ◽  
Author(s):  
D C Gadsby ◽  
M Nakao
Keyword(s):  
Steady State ◽  
Guinea Pig ◽  
Rate Constants ◽  
Ventricular Myocytes ◽  
Pump Current ◽  
Membrane Current ◽  
Negative Slope ◽  
Current Voltage ◽  
State Cycle ◽  
External K

Whole-cell currents were recorded in guinea pig ventricular myocytes at approximately 36 degrees C before, during, and after exposure to maximally effective concentrations of strophanthidin, a cardiotonic steroid and specific inhibitor of the Na/K pump. Wide-tipped pipettes, in combination with a device for exchanging the solution inside the pipette, afforded reasonable control of the ionic composition of the intracellular solution and of the membrane potential. Internal and external solutions were designed to minimize channel currents and Na/Ca exchange current while sustaining vigorous forward Na/K transport, monitored as strophanthidin-sensitive current. 100-ms voltage pulses from the -40 mV holding potential were used to determine steady-state levels of membrane current between -140 and +60 mV. Control experiments demonstrated that if the Na/K pump cycle were first arrested, e.g., by withdrawal of external K, or of both internal and external Na, then neither strophanthidin nor its vehicle, dimethylsulfoxide, had any discernible effect on steady-state membrane current. Further controls showed that, with the Na/K pump inhibited by strophanthidin, membrane current was insensitive to changes of external [K] between 5.4 and 0 mM and was little altered by changing the pipette [Na] from 0 to 50 mM. Strophanthidin-sensitive current therefore closely approximated Na/K pump current, and was virtually free of contamination by current components altered by the changes in extracellular [K] and intracellular [Na] expected to accompany pump inhibition. The steady-state Na/K pump current-voltage (I-V) relationship, with the pump strongly activated by 5.4 mM external K and 50 mM internal Na (and 10 mM ATP), was sigmoid in shape with a steep positive slope between about 0 and -100 mV, a less steep slope at more negative potentials, and an extremely shallow slope at positive potentials; no region of negative slope was found. That shape of I-V relationship can be generated by a two-state cycle with one pair of voltage-sensitive rate constants and one pair of voltage-insensitive rate constants: such a two-state scheme is a valid steady-state representation of a multi-state cycle that includes only a single voltage-sensitive step.

Effects of sevoflurane on inward rectifier K+ current in guinea pig ventricular cardiomyocytes

1997 ◽  
Vol 273 (1) ◽  
pp. H324-H332 ◽  
Author(s):  
A. Stadnicka ◽  
Z. J. Bosnjak ◽  
J. P. Kampine ◽  
W. M. Kwok
Keyword(s):  
Steady State ◽  
Guinea Pig ◽  
Outward Current ◽  
Inward Rectifier ◽  
Equilibrium Potential ◽  
Ventricular Cardiomyocytes ◽  
Steady State Current ◽  
Voltage Dependent ◽  
Current Activation ◽  
Extracellular Side

The effects of sevoflurane on the inward rectifier potassium current (IKIR) were examined in guinea pig ventricular cardiomyocytes using the whole cell patch-clamp methodology. Sevoflurane had a unique dual effect on the steady-state current amplitude, producing a reversible, concentration- and voltage-dependent block of the inward current at potentials negative to the potassium equilibrium potential (EK) but enhancing the outward current positive to EK. Accordingly, the steady-state conductance negative to EK was reduced by sevoflurane, but conductance positive to EK was increased. The chord conductance-voltage relationship showed depolarizing shifts at 0.7, 1.3, and 1.6 mM sevoflurane. When the myocytes were dialyzed with 10 mM Mg2+, but not with 1.0 mM Mg2+, sevoflurane further slowed current activation kinetics. With 10 mM intracellular Mg2+, the outward current enhancement by sevoflurane and the associated shifts in half-activation potential were abolished. Polyamines abolished all effects of sevoflurane on IKIR. With the use of the Woodhull model for voltage-dependent block, we determined the sevoflurane interaction site with the inward rectifier potassium channel to be at an electrical distance of 0.2 from the extracellular side.

scholarly journals Cat Heart Muscle in Vitro

1962 ◽  
Vol 46 (2) ◽  
pp. 189-199 ◽  
Author(s):  
Ernest Page
Keyword(s):  
Steady State ◽  
Heart Muscle ◽  
Potential Difference ◽  
Resting Potential ◽  
Equilibrium Potential ◽  
Papillary Muscles ◽  
K Concentration ◽  
Cat Heart ◽  
External K

The steady state transmembrane resting potential difference (Vm) has been measured in quiescent papillary muscles. Vm was determined as a function of the external K concentration in Cl and SO4 solutions and compared with the K equilibrium potential. Other measurements were made after replacement of external Na by choline, K by Rb and Cs, and Cl by SO4, CH3SO4, and NO3. Effects on Vm of albumin, temperature, and variation in internal K concentration are described.

scholarly journals Voltage-dependent block of cardiac inward-rectifying potassium current by monovalent cations.

1989 ◽  
Vol 94 (2) ◽  
pp. 349-361 ◽  
Author(s):  
R D Harvey ◽  
R E Ten Eick
Keyword(s):  
Ventricular Myocytes ◽  
Negative Slope ◽  
Voltage Sensitivity ◽  
Dependent Manner ◽  
Inward Current ◽  
Current Voltage ◽  
Steady State Current ◽  
Voltage Dependent ◽  
Current Block ◽  
K Current

The inward-rectifying K+ current (IK1) in cat ventricular myocytes, like inward-rectifying K+ currents in many other preparations, exhibited a negative slope conductance region at hyperpolarized membrane potentials that was time-dependent. This was evident as an inactivation of inward current elicited by hyperpolarizing voltage-clamp pulses resulting in a negative slope region of the steady-state current-voltage relationship at potentials negative to -140 mV. Removing extracellular Na+ prevented the development of the negative slope in this voltage region, suggesting that Na+ can block IK1 channels in a time- and voltage-dependent manner. The time and voltage dependence of Cs+-induced block of IK1 was also examined. Cs+ blocked inward current in a manner similar to that of Na+, but the former was much more potent. The fraction of current blocked by Cs+ in the presence of Na+ was reduced in a time- and voltage-dependent manner, which suggested that these blocking ions compete for a common or at least similar site of action. In the absence of Na+, inactivation of IK1 could also be induced by both Cs+ and Li+. However, Li+ was less potent than Na+ in this respect. Calculation of the voltage sensitivity of current block by each of these ions suggests that the mechanism of block by each is similar.

scholarly journals Cation permeation through the voltage-dependent potassium channel in the squid axon. Characteristics and mechanisms.

1987 ◽  
Vol 90 (2) ◽  
pp. 261-290 ◽  
Author(s):  
P K Wagoner ◽  
G S Oxford
Keyword(s):  
Binding Energies ◽  
Ion Concentration ◽  
K Channel ◽  
Delayed Rectifier ◽  
Dependent Manner ◽  
K Channels ◽  
Energy Profiles ◽  
Current Voltage ◽  
Voltage Dependent ◽  
Ion Binding Sites

Characteristics of cation permeation through voltage-dependent delayed rectifier K channels in squid giant axons were examined. Axial wire voltage-clamp measurements and internal perfusion were used to determine conductance and permeability properties. These K channels exhibit conductance saturation and decline with increases in symmetrical K+ concentrations to 3 M. They also produce ion- and concentration-dependent current-voltage shapes. K channel permeability ratios obtained with substitutions of internal Rb+ or NH+4 for K+ are higher than for external substitution of these ions. Furthermore, conductance and permeability ratios of NH+4 or Rb+ to K+ are functions of ion concentration. Conductance measurements also reveal the presence of an anomalous mole fraction effect for NH+4, Rb+, or Tl+ to K+. Finally, internal Cs+ blocks these K channels in a voltage-dependent manner, with relief of block by elevations in external K+ but not external NH+4 or Cs+. Energy profiles for K+, NH+4, Rb+, Tl+, and Cs+ incorporating three barriers and two ion-binding sites are fitted to the data. The profiles are asymmetric with respect to the center of the electric field, have different binding energies and electrical positions for each ion, and (for K+) exhibit concentration-dependent barrier positions.

scholarly journals The Ventral Photoreceptor Cells of Limulus

1969 ◽  
Vol 54 (3) ◽  
pp. 331-351 ◽  
Author(s):  
Ronald Millecchia ◽  
Alexander Mauro
Keyword(s):  
Steady State ◽  
Voltage Clamp ◽  
Dark Current ◽  
Time Course ◽  
Reversal Potential ◽  
Negative Slope ◽  
Induced Current ◽  
Current Voltage ◽  
Steady State Current ◽  
Current Voltage Relation

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.

scholarly journals Conductive Properties and Gating of Channels Formed by Syringopeptin 25A, a Bioactive Lipodepsipeptide from Pseudomonas syringae pv. syringae, in Planar Lipid Membranes

1999 ◽  
Vol 12 (5) ◽  
pp. 401-409 ◽  
Author(s):  
Mauro Dalla Serra ◽  
Ivonne Bernhart ◽  
Paola Nordera ◽  
Domenico Di Giorgio ◽  
Alessandro Ballio ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Lipid Membranes ◽  
Pore Radius ◽  
Maximal Conductance ◽  
Channel Opening ◽  
Current Voltage ◽  
Voltage Dependent ◽  
Planar Lipid Membranes ◽  
Opening And Closing ◽  
Conductive Properties

Syringopeptin 25A, a pseudomonad lipodepsipeptide, can form ion channels in planar lipid membranes. Pore conductance is around 40 pS in 0.1 M NaCl. Channel opening is strongly voltage dependent and requires a negative potential on the same side of the membrane where the toxin was added. These pores open and close with a lifetime of several seconds. At negative voltages, an additional pore state of around 10 pS and a lifetime of around 30 ms is also present. The voltage dependence of the rates of opening and closing of the stable pores is exponential. This allows estimation of the equivalent charge that is moved across the membrane during the process of opening at about 2.6 elementary charges. When NaCl is present, the pore is roughly 3 times more permeant for anions than for cations. The current voltage characteristic of the pore is nonlinear, i.e., pore conductance is larger at negative than at positive voltages. The maximal conductance of the pore depends on the concentration of the salt present, in a way that varies almost linearly with the conductivity of the solution. From this, an estimate of a minimal pore radius of 0.4 nm was derived.

scholarly journals Effects of magnesium on inactivation of the voltage-gated calcium current in cardiac myocytes.

1989 ◽  
Vol 94 (4) ◽  
pp. 745-767 ◽  
Author(s):  
H C Hartzell ◽  
R E White
Keyword(s):  
Steady State ◽  
Ventricular Myocytes ◽  
Internal Surface ◽  
State Level ◽  
Inactivation Kinetics ◽  
Ca Channel ◽  
Patch Clamp Technique ◽  
Inactivation Curve ◽  
Current Voltage ◽  
Voltage Dependent

The effects of changes in intracellular and extracellular free ionized [Mg2+] on inactivation of ICa and IBa in isolated ventricular myocytes of the frog were investigated using the whole-cell configuration of the patch-clamp technique. Intracellular [Mg2+] was varied by internal perfusion with solutions having different calculated free [Mg2+]. Increasing [Mg2+]i from 0.3 mM to 3.0 mM caused a 16% reduction in peak ICa amplitude and a 36% reduction in peak IBa amplitude, shifted the current-voltage relationship and the inactivation curve approximately 10 mV to the left, decreased relief from inactivation, and caused a dramatic increase in the rate of inactivation of IBa. The shifts in the current-voltage and inactivation curves were attributed to screening of internal surface charge by Mg2+. The increased rate of inactivation of IBa was due to an increase in both the steady-state level of inactivation as well as an increase in the rate of inactivation, as measured by two-pulse inactivation protocols. Increasing external [Mg2+] decreased IBa amplitude and shifted the current-voltage and inactivation curves to the right, but, in contrast to the effect of internal Mg2+, had little effect on the inactivation kinetics or the steady-state inactivation of IBa at potentials positive to 0 mV. These observations suggest that the Ca channel can be blocked quite rapidly by external Mg2+, whereas the block by [Mg2+]i is time and voltage dependent. We propose that inactivation of Ca channels can occur by both calcium-dependent and purely voltage-dependent mechanisms, and that a component of voltage-dependent inactivation can be modulated by changes in cytoplasmic Mg2+.

Potential-dependent block of human delayed rectifier K+ channels by internal Na+

1996 ◽  
Vol 270 (4) ◽  
pp. C1131-C1144 ◽  
Author(s):  
S. Johansson ◽  
A. K. Sundgren ◽  
U. Kahl
Keyword(s):  
Single Channel ◽  
Neuroblastoma Cells ◽  
Negative Slope ◽  
Delayed Rectifier ◽  
Na Current ◽  
Intact Cells ◽  
K Channels ◽  
High Potentials ◽  
K Current ◽  
K Currents

The delayed rectifier K+ currents in differentiated human SH-SY5Y neuroblastoma cells were characterized with tight-seal recording techniques. Activation and inactivation parameters were measured. At high positive potentials, the current showed a marked rectification, causing a region of negative slope conductance in the current vs. potential curve. The rectification depended markedly on the pipette Na+ concentration. Without Na+, no rectification was observed, whereas with high Na+ (20-60 mM), a marked rectification was always observed. Tail current measurements showed a fast ( < 400 microseconds) block of K+ currents in the presence of internal Na+. With 60 mM Na+ in the pipette 8% of the K+ current was blocked at 0 mV, 27% at +20 mV, and 82% at +100 mV. Similar degrees of block were often seen with 30 mM Na+ in the pipette. The submembrane Na+ concentration in intact cells was estimated, on the basis of the reversal of Na+ current, to be approximately 15 mM. Single-channel K+ currents, in the cell-attached configuration, showed a conductance of approximately 20 pS at 40-60 mV above rest but showed rectification at high potentials.

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