scholarly journals Potassium flux ratio in voltage-clamped squid giant axons.

1980 ◽  
Vol 76 (1) ◽  
pp. 83-98 ◽  
Author(s):  
T Begenisich ◽  
P De Weer
Keyword(s):  
Simultaneous Measurement ◽  
Potassium Conductance ◽  
Flux Ratio ◽  
Giant Axon ◽  
Equilibrium Potential ◽  
Giant Axons ◽  
Average Value ◽  
Potassium Flux ◽  
K Current ◽  
Loligo Pealei

The potassium flux ratio across the axolemma of internally perfused, voltage-clamped giant axons of Loligo pealei has been evaluated at various membrane potentials and internal potassium concentrations ([K]i). Four different methods were used: (a) independent measurement of one-way influx and efflux of 42K; (b) simultaneous measurement of net K current (IK) and 42K influx; (c) simultaneous measurement of IK and 42K efflux; and (d) measurement of potassium conductance and 42K influx at the potassium equilibrium potential. The reliability of each of these methods is discussed. The average value of the exponent n' in the Hodgkin-Keynes equation ranged from 1.5 at -4mV and 200 mM [K]i to 3.3 at -38 mV and 350 mM [K]i and appeared to be a function of membrane potential and possibly of [K]i. It is concluded that the potassium channel of squid giant axon is a multi-ion, single-file pore with three or more sites.

scholarly journals Blockage of Sodium Conductance Increase in Lobster Giant Axon by Tarichatoxin (Tetrodotoxin)

1966 ◽  
Vol 49 (5) ◽  
pp. 977-988 ◽  
Author(s):  
M. Takata ◽  
J. W. Moore ◽  
C. Y. Kao ◽  
F. A. Fuhrman
Keyword(s):  
Potassium Conductance ◽  
Giant Axon ◽  
High Concentration ◽  
Temporal Characteristics ◽  
Giant Axons ◽  
Sodium Conductance ◽  
Puffer Fish ◽  
Conductance Increase

Tarichatoxin, isolated from California newt eggs, has been found to selectively block the increase of sodium conductance associated with excitation in lobster giant axons at nanomolar concentrations. This resulted from a reduction in the amplitude of the conductance increase rather than a change in its temporal characteristics. The normal potassium conductance increase with depolarization is not altered. A high concentration of calcium applied concomitantly with the toxin significantly improves the reversibility of the sodium blocking. This toxin has recently been identified as chemically identical with tetrodotoxin from the puffer fish. Toxins from the two sources are equally effective and are shown to have an action which is distinctly different from that of procaine.

scholarly journals Sodium Flux Ratio in Na/K Pump-Channels Opened by Palytoxin

2007 ◽  
Vol 130 (1) ◽  
pp. 41-54 ◽  
Author(s):  
R.F. Rakowski ◽  
Pablo Artigas ◽  
Francisco Palma ◽  
Miguel Holmgren ◽  
Paul De Weer ◽  
...  
Keyword(s):  
Binding Sites ◽  
Reversal Potential ◽  
Flux Ratio ◽  
Giant Fiber ◽  
Animal Cells ◽  
Giant Axons ◽  
Single File ◽  
Sodium Flux ◽  
Na Ions ◽  
Loligo Pealei

Palytoxin binds to Na+/K+ pumps in the plasma membrane of animal cells and opens an electrodiffusive cation pathway through the pumps. We investigated properties of the palytoxin-opened channels by recording macroscopic and microscopic currents in cell bodies of neurons from the giant fiber lobe, and by simultaneously measuring net current and 22Na+ efflux in voltage-clamped, internally dialyzed giant axons of the squid Loligo pealei. The conductance of single palytoxin-bound “pump-channels” in outside-out patches was ∼7 pS in symmetrical 500 mM [Na+], comparable to findings in other cells. In these high-[Na+], K+-free solutions, with 5 mM cytoplasmic [ATP], the K0.5 for palytoxin action was ∼70 pM. The pump-channels were ∼40–50 times less permeable to N-methyl-d-glucamine (NMG+) than to Na+. The reversal potential of palytoxin-elicited current under biionic conditions, with the same concentration of a different permeant cation on each side of the membrane, was independent of the concentration of those ions over the range 55–550 mM. In giant axons, the Ussing flux ratio exponent (n') for Na+ movements through palytoxin-bound pump-channels, over a 100–400 mM range of external [Na+] and 0 to −40 mV range of membrane potentials, averaged 1.05 ± 0.02 (n = 28). These findings are consistent with occupancy of palytoxin-bound Na+/K+ pump-channels either by a single Na+ ion or by two Na+ ions as might be anticipated from other work; idiosyncratic constraints are needed if the two Na+ ions occupy a single-file pore, but not if they occupy side-by-side binding sites, as observed in related structures, and if only one of the sites is readily accessible from both sides of the membrane.

Excitability of the Squid Giant Axon Revisited

1998 ◽  
Vol 80 (2) ◽  
pp. 903-913 ◽  
Author(s):  
John R. Clay
Keyword(s):  
Standard Model ◽  
Action Potential ◽  
Giant Axon ◽  
Squid Giant Axon ◽  
Equilibrium Potential ◽  
Single Action ◽  
Single Action Potential ◽  
Nerve Excitability ◽  
The Standard Model ◽  
Loligo Pealei

Clay, John R. Excitability of the squid giant axon revisited. J. Neurophysiol. 80: 903–913, 1998. The electrical properties of the giant axon from the common squid Loligo pealei have been reexamined. The primary motivation for this work was the observation that the refractoriness of the axon was significantly greater than the predictions of the standard model of nerve excitability. In particular, the axon fired only once in response to a sustained, suprathreshold stimulus. Similarly, only a single action potential was observed in response to the first pulse of a train of 1-ms duration current pulses, when the pulses were separated in time by ∼10 ms. The axon was refractory to all subsequent pulses in the train. The underlying mechanisms for these results concern both the sodium and potassium ion currents I Na and I K. Specifically, Na+ channel activation has long been known to be coupled to inactivation during a depolarizing voltage-clamp step. This feature appears to be required to simulate the pulse train results in a revised model of nerve excitability. Moreover, the activation curve for I K has a significantly steeper voltage dependence, especially near its threshold (approximately −60 mV), than in the standard model, which contributes to reduced excitability, and the fully activated current-voltage relation for I K has a nonlinear, rather than a linear, dependence on driving force. An additional aspect of the revised model is accumulation/depeletion of K+ in the space between the axon and the glial cells surrounding the axon, which is significant even during a single action potential and which can account for the 15–20 mV difference between the potassium equilibrium potential E K and the maximum afterhyperpolarization of the action potential. The modifications in I K can also account for the shape of voltage changes near the foot of the action potential.

Carbachol induces oscillations of membrane potassium conductance in a colonic cell line, T84

1990 ◽  
Vol 258 (2) ◽  
pp. C318-C326 ◽  
Author(s):  
D. C. Devor ◽  
S. M. Simasko ◽  
M. E. Duffey
Keyword(s):  
Membrane Potential ◽  
Potassium Conductance ◽  
Equilibrium Potential ◽  
Base Line ◽  
Resting Cells ◽  
T84 Cells ◽  
Potential Oscillations ◽  
Pipette Solution ◽  
K Current ◽  
Cl Conductance

Effects of carbachol on membrane potential and current in T84 cells were determined using whole cell patch-clamp techniques. When the pipettes contained a standard KCl solution and the bath contained a standard NaCl solution, carbachol (100 microM) caused a rapid hyperpolarization to the K+ equilibrium potential (EK+), followed by potential oscillations. When membrane potential was clamped to 0 mV, carbachol induced an outwardly directed K+ current in 31 of 37 cells, with a peak value of 618 +/- 51 (SE) pA. In 77% of these cells the current oscillated and gradually declined to base line. Atropine (20 microM) blocked this response. In symmetric KCl solutions the carbachol-induced current reversed at 0 mV with no rectification. Ba2+ or Cs+ did not block the current, but tetraethylammonium ion (TEA) reduced the number of responding cells. Although a Cl- conductance was found in resting cells, carbachol did not cause an increase in Cl- current when the cells were voltage-clamped to EK+, or when voltage-clamped to +/- 60 mV while bathed in symmetric NaCl solutions. When the Ca2(+)-buffering capacity of the pipette solution was increased, 80% of the cells responded to carbachol, but only 10% oscillated; however, no K+ current was induced by carbachol when the pipette was made nominally Ca2+ free. The current was not affected by removal of Ca2+ from the bath. These results show that carbachol induces an oscillating Ca2(+)-activated K+ conductance in T84 cells, but no Cl- conductance. This K+ conductance is dependent on the mechanisms that regulate intracellular Ca2+.

scholarly journals Sodium flux ratio in voltage-clamped squid giant axons.

1981 ◽  
Vol 77 (5) ◽  
pp. 489-502 ◽  
Author(s):  
T Begenisich ◽  
D Busath
Keyword(s):  
Sodium Channels ◽  
Binding Sites ◽  
Flux Ratio ◽  
Ion Binding ◽  
Ion Permeation ◽  
Giant Axons ◽  
Sodium Flux ◽  
Ion Binding Sites ◽  
Loligo Pealei ◽  
Ion Pore

The sodium flux ratio across the axolemma of internally perfused, voltage-clamped giant axons of Loligo pealei has been measured at various membrane potentials. The flux ratio exponent obtained from these measurements was about unity and independent of membrane voltage over the 50 mV range from about -20 to +30 mV. These results, combined with previous measurements of ion permeation through sodium channels, show that the sodium channel behaves like a multi-ion pore with two ion binding sites that are rarely simultaneously occupied by sodium.

Preliminary observations on escape swimming and giant neurons in Aglantha digitale (Hydromedusae: Trachylina)

1980 ◽  
Vol 58 (4) ◽  
pp. 549-552 ◽  
Author(s):  
S. Donaldson ◽  
G. O. Mackie ◽  
A. Roberts
Keyword(s):  
Action Potentials ◽  
Giant Axon ◽  
Synaptic Delay ◽  
Giant Axons ◽  
A Value ◽  
Giant Synapses ◽  
Run Up

Aglantha can swim in two ways, one of which, fast swimming, is evoked by contact with predators and serves for escape. The response consists of two or three violent contractions of which the first propels the animal a distance equivalent to five body lengths. Peak velocities in the range 0.3–0.4 m s−1 were measured. Drag is reduced by contraction of the tentacles.Coordination of escape swimming and tentacle contraction is achieved by a system of giant axons. A giant axon runs down each tentacle; action potentials in these elements show a one-for-one correspondence with potentials recorded from a ring-shaped axon lying in the margin near the tentacle bases. The ring giant synapses with eight motor giants which run up the subumbrella innervating the swimming muscles.Conduction velocities in the giant axons may be as high as 4.0 m s−1 in the case of the largest (40 μm diameter) axons. A value of 1.6 ms was obtained for minimum synaptic delay between the ring and motor giant axons.

scholarly journals Activation-inactivation of potassium channels and development of the potassium-channel spike in internally perfused squid giant axons.

1981 ◽  
Vol 78 (1) ◽  
pp. 43-61 ◽  
Author(s):  
I Inoue
Keyword(s):  
Potassium Channels ◽  
Potassium Channel ◽  
Voltage Clamp ◽  
Time Constant ◽  
Equilibrium Potential ◽  
Giant Axons ◽  
Dependent Inactivation ◽  
Voltage Dependent ◽  
Time Courses ◽  
Calcium Permeability

A spike that is the result of calcium permeability through potassium channels was separated from the action potential is squid giant axons internally perfused with a 30 mM NaF solution and bathed in a 100 mM CaCl2 solution by blocking sodium channels with tetrodotoxin. Currents through potassium channels were studied under voltage clamp. The records showed a clear voltage-dependent inactivation of the currents. The inactivation was composed of at least two components; one relatively fast, having a time constant of 20--30 ms, and the other very slow, having a time constant of 5--10 s. Voltage clamp was carried out with a variety of salt compositions in both the internal and external solutions. A similar voltage-dependent inactivation, also composed of the two components, was recognized in all the current through potassium channels. Although the direction and intensity of current strongly depended on the salt composition of the solutions, the time-courses of these currents at corresponding voltages were very similar. These results strongly suggest that the inactivation of the currents in attributable to an essential, dynamic property of potassium channels themselves. Thus, the generation of a potassium-channel spike can be understood as an event that occurs when the equilibrium potential across the potassium channel becomes positive.

Coordinated Excitation of Flexor Inhibitors in the Crayfish

1980 ◽  
Vol 86 (1) ◽  
pp. 187-195
Author(s):  
CHIKAO UYAMA ◽  
TAKASHI MATSUYAMA
Keyword(s):  
Giant Axon ◽  
Experimental Results ◽  
Abdominal Ganglion ◽  
Giant Axons ◽  
The Third ◽  
Nerve Cords

In isolated abdominal nerve cords of crayfish, the medial or lateral giant axons were stimulated at a position just rostral to the first abdominal ganglion. Recordings of the impulse sequences of the flexor inhibitor (FI) were made from the anterior five ganglia, three ganglia at a time. In 20% of our preparations, one giant axon impulse caused one to four FI impulses in every abdominal third root. An equal number of FI impulses were usually produced by each abdominal ganglion for any given stimulation. The earliest FI impulse was observed at the third root of the fourth ganglion. FI impulses occurred with increasing latencies rostrally and caudally from the fourth ganglion. The FI responses to medial and lateral giant axons stimulation were essentially equivalent. FI impulses were recorded from the rostral three abdominal ganglia, while the caudal ganglia were cut off one after another from the sixth to the third ganglion. Little change was noted until after the removal of the fourth ganglion, which usually caused all FI impulses to disappear. From these experimental results, we propose a model of central mechanisms for FI excitation.

Neuromuscular transmission in the jellyfish Aglantha digitale

1985 ◽  
Vol 116 (1) ◽  
pp. 1-25 ◽  
Author(s):  
P. A. Kerfoot ◽  
G. O. Mackie ◽  
R. W. Meech ◽  
A. Roberts ◽  
C. L. Singla
Keyword(s):  
Dimensional Space ◽  
Thin Sheet ◽  
Current Source ◽  
Giant Axon ◽  
The Body ◽  
Two Dimensional ◽  
Bathing Medium ◽  
Giant Axons ◽  
Motor Neurones ◽  
Stimulation Of

In the jellyfish Aglantha digitale escape swimming is mediated by the nearly synchronous activity of eight giant motor axons which make direct synaptic contact with contractile myoepithelial cells on the under-surface of the body wall. The delay in transmission at these synapses was 0.7 +/− 0.1 ms (+/− S.D.;N = 6) at 12 degrees C as measured from intracellular records. Transmission depended on the presence of Ca2+ in the bathing medium. It was not blocked by increasing the level of Mg2+ to 127 mmol l-1. The myoepithelium is a thin sheet of electrically coupled cells and injection of current at one point was found to depolarize the surrounding cells. The potential change declined with distance from the current source as expected for two-dimensional current spread. The two-dimensional space constant (lambda) was 770 micron for current flow in the circular direction and 177 micron for radial flow. The internal resistance of the epithelium (178–201 omega cm) and the membrane time constant (5–10 ms) were direction independent. No propagated epithelial action potentials were observed. Spontaneous miniature synaptic potentials of similar amplitude and rise-time were recorded intracellularly at distances of up to 1 mm from the motor giant axon. Ultrastructural evidence confirms that neuro-myoepithelial synapses also occur away from the giant axons. It is likely that synaptic sites are widespread in the myoepithelium, probably associated with the lateral motor neurones as well as the giant axons. Local stimulation of lateral motor neurones generally produced contraction in distinct fields. We suppose that stimulation of a single motor giant axon excites a whole population of lateral motor neurones and hence a broad area of the myoepithelium.

G proteins activate ionic conductances at multiple sites in T84 cells

1997 ◽  
Vol 272 (4) ◽  
pp. C1222-C1231
Author(s):  
L. Izu ◽  
M. Li ◽  
R. DeMuro ◽  
M. E. Duffey
Keyword(s):  
G Protein ◽  
G Proteins ◽  
Cell Voltage ◽  
Equilibrium Potential ◽  
T84 Cells ◽  
Ionic Conductances ◽  
Inward Currents ◽  
Cl Channels ◽  
K Current

We examined the role of G proteins in activation of ionic conductances in isolated T84 cells during cholinergic stimulation. When cells were whole cell voltage clamped to the K+ equilibrium potential (E(K)) or Cl- equilibrium potential (E(Cl)) under standard conditions, the cholinergic agonist, carbachol, induced a large oscillating K+ current but only a small inward current. Addition of the GDP analogue, guanosine 5'-O-(2-thiodiphosphate), to pipettes blocked the ability of carbachol to activate the K+ current. Addition of the nonhydrolyzable GTP analogue, guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS), to pipettes stimulated large oscillating K+ and inward currents. This occurred even when Ca2+ was absent from the bath but not when the Ca2+ chelator, ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid, was added to pipettes. When all pipette and bath K+ was replaced with Na+ and cells were voltage clamped between E(Na) and E(Cl), GTPgammaS activated oscillating Na+ and Cl- currents. Finally, addition of inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] to pipettes activated large oscillating K+ currents but only small inward currents. These results suggest that a carbachol-induced release of Ca2+ from intracellular stores is activated by a G protein through the phospholipase C-Ins(1,4,5)P3 signaling pathway. In addition, this or another G protein activates Cl- current by directly gating Cl- channels to increase their sensitivity to Ca2+.

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