scholarly journals Effects of Batrachotoxin on Membrane Potential and Conductance of Squid Giant Axons

1971 ◽  
Vol 58 (1) ◽  
pp. 54-70 ◽  
Author(s):  
Toshio Narahashi ◽  
Edson X. Albuquerque ◽  
Takehiko Deguchi

The effects of batrachotoxin (BTX) on the membrane potential and conductances of squid giant axons have been studied by means of intracellular microelectrode recording, internal perfusion, and voltage clamp techniques. BTX (550–1100 nM) caused a marked and irreversible depolarization of the nerve membrane, the membrane potential being eventually reversed in polarity by as much as 15 mv. The depolarization progressed more rapidly with internal application than with external application of BTX to the axon. External application of tetrodotoxin (1000 nM) completely restored the BTX depolarization. Removal or drastic reduction of external sodium caused a hyperpolarization of the BTX-poisoned membrane. However, no change in the resting membrane potential occurred when BTX was applied in the absence of sodium ions in both external and internal phases. These observations demonstrate that BTX specifically increases the resting sodium permeability of the squid axon membrane. Despite such an increase in resting sodium permeability, the BTX-poisoned membrane was still capable of undergoing a large sodium permeability increase of normal magnitude upon depolarizing stimulation provided that the membrane potential was brought back to the original or higher level. The possibility that a single sodium channel is operative for both the resting sodium, permeability and the sodium permeability increase upon stimulation is discussed.

1976 ◽  
Vol 67 (3) ◽  
pp. 369-380 ◽  
Author(s):  
J Villegas ◽  
C Sevcik ◽  
F V Barnola ◽  
R Villegas

The actions of grayanotoxin I, veratrine, and tetrodotoxin on the membrane potential of the Schwann cell were studied in the giant nerve fiber of the squid Sepioteuthis sepioidea. Schwann cells of intact nerve fibers and Schwann cells attached to axons cut lengthwise over several millimeters were utilized. The axon membrane potential in the intact nerve fibers was also monitored. The effects of grayanotoxin I and veratrine on the membrane potential of the Schwann cell were found to be similar to those they produce on the resting membrane potential of the giant axon. Thus, grayanotoxin I (1-30 muM) and veratrine (5-50 mug-jl-1), externally applied to the intact nerve fiber or to axon-free nerve fiber sheaths, produce a Schwann cell depolarization which can be reversed by decreasing the external sodium concentration or by external application of tetrodotoxin. The magnitude of these membrane potential changes is related to the concentrations of the drugs in the external medium. These results indicate the existence of sodium pathways in the electrically unexcitable Schwann cell membrane of S. sepioidea, which can be opened up by grayanotoxin I and veratrine, and afterwards are blocked by tetrodotoxin. The sodium pathways of the Schwann cell membrane appear to be different from those of the axolemma which show a voltage-dependent conductance.


1978 ◽  
Vol 75 (1) ◽  
pp. 253-263
Author(s):  
J. E. TREHERNE ◽  
Y. PICHON

Reprint requests should be addressed to Dr Treherne. Sabella is a euryhaline osmoconformer which is killed by direct transfer to 50% sea water, but can adapt to this salinity with progressive dilution of the sea water. The giant axons were adapted to progressive dilution of the bathing medium (both in vivo and in vitro) and were able to function at hyposmotic dilutions (down to 50%) sufficient to induce conduction block in unadapted axons. Hyposmotic adaptation of the giant axon involves a decrease in intracellular potassium concentration which tends to maintain a relatively constant resting potential during adaptation despite the reduction in external potassium concentration. There is no appreciable change in the intracellular sodium concentration, but the relative sodium permeability of the active membrane increases during hyposmotic adaptation. This increase partially compensates for the reduction in sodium gradient across the axon membrane, during dilution of the bathing media, by increasing the overshoot of the action potentials recorded in hyposmotically adapted axons.


1982 ◽  
Vol 242 (5) ◽  
pp. C296-C303 ◽  
Author(s):  
B. Ribalet ◽  
P. M. Beigelman

The present studies, designed to evaluate the contribution of Na+ to the mouse pancreatic beta-cell membrane potential, were performed utilizing intracellular microelectrodes. Complete removal of external sodium, in the presence of glucose, did not significantly affect spike peak potential. However, it caused a negative shift of the resting membrane potential, both in the presence and absence of glucose. After this initial hyperpolarization, the membrane gradually depolarized, the rate of depolarization being slower in the absence of glucose. This two-phase hyperpolarization-depolarization pattern remained when ouabain was added, both in the presence and absence of glucose. An increase of input resistance was associated with the slow depolarization. During this depolarization the maximum rate of rise (dV/dtmax) of the action potential (“spike”) decreased. There was no direct relationship between dV/dtmax and [Na]0. Readdition of low [Na]0 (14 mM) to a glucose medium reactivated the postburst hyperpolarization (PBH), even in the presence of ouabain. These observations indicate that there is a significant resting sodium permeability (PNa). However, the action potential (spike) is not generated by activation of a voltage-dependent (gated) sodium channel. The membrane depolarization after Na+ removal reflects concomitant inhibition of the Na+-K+ pump and decrease of potassium permeability (PK). The blockage of PBH in the absence of Na+ is not related to the inhibition of an oscillatory Na+-K+ pump but to the inactivation of a PK. Aside from its effect on the Na+-K+ pump, ouabain may stimulate PNa.


1941 ◽  
Vol 25 (1) ◽  
pp. 29-51 ◽  
Author(s):  
Kenneth S. Cole

Previous measurements have shown that the electrical properties of the squid axon membrane are approximately equivalent to those of a circuit containing a capacity shunted by an inductance and a rectifier in series. Selective ion permeability of a membrane separating two electrolytes may be expected to give rise to the rectification. A quasi-crystalline piezoelectric structure of the membrane is a plausible explanation of the inductance. Some approximate calculations of behavior of an axon with these membrane characteristics have been made. Fair agreement is obtained with the observed constant current subthreshold potential and impedance during the foot of the action potential. In a simple case a formal analogy is found between the calculated membrane potential and the excitability defined by the two factor formulations of excitation. Several excitation phenomena may then be explained semi-quantitatively by further assuming the excitability proportional to the membrane potential. Some previous measurements and subthreshold potential and excitability observations are not consistent with the circuit considered and indicate that this circuit is only approximately equivalent to the membrane.


1969 ◽  
Vol 54 (6) ◽  
pp. 730-740 ◽  
Author(s):  
L. Binstock ◽  
L. Goldman

A new dissection procedure for preparing Myxicola giant axons for observation under voltage clamp is described. Preparation time is generally 40–45 min. 65–70% of the preparations attempted may be brought through the entire procedure, including insertion of the long internal electrode, and support an initial action potential amplitude of 100 mv or greater. Mean values for axon diameter, resting membrane potential, action potential amplitude, maximum peak inward transient current, and resting membrane resistance are 560 µ, —66.5 mv, 112 mv, 0.87 ma/cm2 and 1.22 KΩ cm 2 respectively. Cut branches do not seem to be a problem in this preparation. Behavior under voltage clamp is reasonably stable over several hours. Reductions in maximum inward transient current of 10% and in steady-state current of 5–10% are expected in the absence of any particular treatment. Tetrodotoxin blocks the action potential and both the inward and outward transient current, but has no effect on either the resting membrane potential or the steady-state current. This selective action of tetrodotoxin on the transient current is taken as an indication that this current component is probably carried by Na.


1959 ◽  
Vol 196 (5) ◽  
pp. 1071-1078 ◽  
Author(s):  
Robert E. Taylor

Procaine (0.025–0.1%; pH 7.9) caused a reduction in the amount and rate of development of the early transient (sodium) and late steady state (potassium) currents which occur during a depolarizing voltage step applied to the excised, voltage clamped squid axon. Consistent results were obtained by holding the membrane potential at a hyperpolarized value prior to the applied step. No effect was seen on the resting potential, on the sodium equilibrium potential, or on the proportion of the sodium carrying system which was ‘inactive’ at any membrane potential. The blocking action of procaine is a result of the inhibition by the drug of the sodium carrying system. The effect of procaine on the potassium conductance is such as to oppose the blocking action.


1967 ◽  
Vol 50 (7) ◽  
pp. 1835-1848 ◽  
Author(s):  
Joseph P. Senft

The resting membrane potential of the lobster axon becomes 5–8 mv more negative when the temperature of the perfusion solution is increased 10°C. This potential change is about twice that predicted if the axon membrane potential followed that expected for a potassium ion electrode potential. When the inhibitors, 2, 4-dinitrophenol, sodium cyanide, and sodium azide, were added separately to the perfusion medium the potential change was reduced to about 1.4 times that predicted for a potassium ion electrode potential. Assays of axons exposed to these inhibitors showed that ATP levels were reduced to about one-fourth that obtained for control axons. Ouabain added to the perfusion medium reduced the potential change to that expected for a potassium ion electrode potential. These results suggest that the resting potential changes with temperature as a result of the activity of an electrogenic ion pump.


1969 ◽  
Vol 54 (6) ◽  
pp. 741-754 ◽  
Author(s):  
L. Goldman ◽  
L. Binstock

The effect of reducing the external sodium concentration, [Na]o, on resting potential, action potential, membrane current, and transient current reversal potential in Myxicola giant axons was studied. Tris chloride was used as a substitute for NaCl. Preliminary experiments were carried out to insure that the effect of Tris substitution could be attributed entirely to the reduction in [Na]o. Both choline and tetramethylammonium chloride were found to have additional effects on the membrane. The transient current is carried largely by Na, while the delayed current seems to be independent of [Na]o. Transient current reversal potential behaves much like a pure Nernst equilibrium potential for sodium. Small deviations from this behavior are consistent with the possibility of some small nonsodium component in the transient current. An exact PNa/PK for the transient current channels could not be computed from these data, but is certainly well greater than unity and possibly quite large. The peak of the action potential varied with [Na]o as expected for a sodium action potential with some substantial potassium permeability at the time of peak. Resting membrane potential is independent of [Na]o. This finding is inconsistent with the view that the resting membrane potential is determined only by the distribution of K and Na, and PNa/PK. It is suggested that PNa/PK's obtained from resting membrane potential-potassium concentration data do not always have the physical meaning generally attributed to them.


1969 ◽  
Vol 54 (6) ◽  
pp. 755-764 ◽  
Author(s):  
L. Goldman ◽  
L. Binstock

Early leak current, i.e. for times similar to the time to peak of the transient current was measured in Myxicola giant axons in the presence of tetrodotoxin. The leak current-voltage relation rectifies, showing more current for strong depolarizing pulses than expected from symmetry around the holding potential. A satisfactory practical approximation for most leak corrections is constant resting conductance. The leak current-voltage curve rectifies less than expected from the constant field equation. These curves cannot be reconstructed by summing the constant field currents for sodium and potassium using a PNa/PK ratio obtained in the usual way, from zero current constant field fits to resting membrane potential data. Nor can they be reconstructed by summing the constant field current for potassium with that for any other single ion. They can be reconstructed, however, by summing the constant field current for potassium with a constant conductance component. It is concluded that the leak current and the resting membrane potential, therefore, are determined by multiple ionic components, at least three and possibly many. Arguments are presented suggesting that ion permeability ratios obtained in the usual way, by fitting the constant field equation to resting membrane potential data should be viewed with skepticism.


1963 ◽  
Vol 204 (3) ◽  
pp. 480-482 ◽  
Author(s):  
Raymond J. Lipicky ◽  
S. H. Bryant

In squid ( Loligo pealii) giant axons, potassium efflux was measured after soak-loading during washout, and membrane potentials were monitored using an intracellular pipette electrode. As external divalent cation is lowered down to 1/10 of normal, there is little change of resting potassium efflux or resting membrane potential (slight hyperpolarization), despite a tendency of the axon to fire repetitively. Below this concentration, efflux rises as much as eight times the normal 265 pmole·cm–2·sec–1 and potential falls (from 55 as far as 32 mv), inexcitability occurring at about 1/100 of normal external divalent cation concentration.


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