scholarly journals Axonal Adaptations to Osmotic and Ionic Stress in an Invertebrate Osmoconformer (Mercierella Enigmatica Fauvel): I. Ultrastructural and Electrophysiological Observations on Axonal Accessibility

1978 ◽  
Vol 76 (1) ◽  
pp. 191-204
Author(s):  
H. LE B. SKAER ◽  
J. E. TREHERNE ◽  
J. A. BENSON

The giant axons in Mercierella are overlaid by narrow glial processes which provide an incomplete covering of the axonal surface. Where more complete covering occurs the intercellular clefts are not sealed by tight junctional complexes. Ionic lanthanum penetrates to the surfaces of axons from sea-water-adapted animals (in normal saline and during initial exposure to hyposmotic saline) and, also, to the surface of hyposmotically adapted axons. A relatively free intercellular access to the axon surfaces is also indicated by the rapid electrical responses of sea-water-adapted axons to hyposmotic dilution and of hyposmotically adapted axons to sodium-deficient saline. The giant axon possesses an unusual ultrastructural specialization: hemidesmosome-like structures (associated with the axon membrane) which are connected to a network of neuronlaments within the axon. Theoretical considerations suggest that these structures could enable the axons to withstand appreciable excesses in intracellular hydrostatic pressure resulting from osmotic imbalance during hyposmotic stress. Note: The Editor reports, with deep regret, the apparent passing of Mercierella enigmatica which it is now proposed becomes Ficopotamus enigmaticus (Fauvel) (ten Hove & Weerdenburg, 1978, Biol. Bull. 154, 96–120).

1976 ◽  
Vol 69 (1) ◽  
pp. 19-28 ◽  
Author(s):  
GM Villegas ◽  
J Villegas

Giant nerve fibers of squid Sepioteuthis sepiodea were incubated for 10 min in artificial sea water (ASW) under control conditions, in the absence of various ions, and in the presence of cardiac glycosides. The nerve fibers were fixed in OsO(4) and embedded in Epon, and structural complexes along the axolemma were studied. These complexes consist of a portion of axolemma exhibiting a three-layered substructure, an undercoating of a dense material (approximately 0.1μm in length and approximately 70-170 A in thickness), and a narrowing to disappearance of the axon-Schwann cell interspace. In the controls, the incidence of complexes per 1,000μm of axon perimeter was about 137. This number decreased to 10-25 percent when magnesium was not present in the incubating media, whatever the calcium concentration (88, 44, or 0 mM). In the presence of magnesium, the number and structural features of the complexes were preserved, though the number decreased to 65 percent when high calcium was simultaneously present. The complexes were also modified and decreased to 26-32 percent by incubating the nerves in solutions having low concentrations of sodium and potassium. The adding of 10(-5) M ouabain or strophanthoside to normal ASW incubating solution decreased them to 20-40 percent. Due to their sensitivity to changes in external ionic concentrations and to the presence of cardiac glycosides, the complexes are proposed to represent the structural correlate of specialized sites for active ion transport, although other factors may be involved.


1962 ◽  
Vol 46 (2) ◽  
pp. 257-266 ◽  
Author(s):  
Rita Guttman ◽  

The effect of temperature on the potential and current thresholds of the squid giant axon membrane was measured with gross external electrodes. A central segment of the axon, 0.8 mm long and in sea water, was isolated by flowing low conductance, isoosmotic sucrose solution on each side; both ends were depolarized in isoosmotic KCl. Measured biphasic square wave currents at five cycles per second were applied between one end of the nerve and the membrane of the central segment. The membrane potential was recorded between the central sea water and the other depolarized end. The recorded potentials are developed only across the membrane impedance. Threshold current values ranged from 3.2 µa at 267deg;C to 1 µa at 7.5°C. Threshold potential values ranged from 50 mv at 26°C to 6 mv at 7.5°C. The mean Q10 of threshold current was 2.3 (SD = 0.2), while the Q10 for threshold potentials was 2.0 (SD = 0.1).


1960 ◽  
Vol 43 (5) ◽  
pp. 971-980 ◽  
Author(s):  
Kenneth S. Cole ◽  
John W. Moore

The potential differences across the squid giant axon membrane, as measured with a series of microcapillary electrodes filled with concentrations of KCl from 0.03 to 3.0 M or sea water, are consistent with a constant membrane potential and the liquid junction potentials calculated by the Henderson equation. The best value for the mobility of an organic univalent ion, such as isethionate, leads to a probably low, but not impossible, axoplasm specific resistance of 1.2 times sea water and to a liquid junction correction of 4 mv. for microelectrodes filled with 3 M KCl. The errors caused by the assumptions of proportional mixing, unity activity coefficients, and a negligible internal fixed charge cannot be estimated but the results suggest that the cumulative effect of them may not be serious.


1938 ◽  
Vol 21 (6) ◽  
pp. 757-765 ◽  
Author(s):  
Howard J. Curtis ◽  
Kenneth S. Cole

The impedance of the excised giant axon from hindmost stellar nerve of Loligo pealii has been measured over the frequency range from 1 to 2500 kilocycles per second. The measurements have been made with the current flow perpendicular to the axis of the axon to permit a relatively simple analysis of the data. It has been found that the axon membrane has a polarization impedance with an average phase angle of 76° and an average capacity of 1.1µf./cm2 at 1 kilocycle. The direct current resistance of the membrane could not be measured, but was greater than 3 ohm cm.2 and the average internal specific resistance was four times that of sea water. There was no detectable change in the membrane impedance when the axon lost excitability, but some time later it decreased to zero.


1962 ◽  
Vol 45 (6) ◽  
pp. 1217-1238 ◽  
Author(s):  
Fred J. Julian ◽  
John W. Moore ◽  
David E. Goldman

The sucrose-gap method introduced by Stämpfli provides a means for the application of a voltage clamp to the lobster giant axon, which responds to a variety of different experimental procedures in ways quite similar to those reported for the squid axon and frog node. This is particularly true for the behavior of the peak initial current. However, the steady state current shows some differences. It has a variable slope conductance less than that of the peak initial current. The magnitude of the steady state slope conductance is related to the length of the repolarization phase of the action potential, which does not have an undershoot in the lobster. The steady state outward current is maintained for as long as 100 msec.; this is in contrast to a decline of about 50 per cent in the squid axon. Lowering the external calcium concentration produces shifts in the current-voltage relations qualitatively similar to those obtained from the squid axon. On the basis of the data available, there is no reason to doubt that the Hodgkin and Huxley analysis for the squid giant axon in sea water can be applied to the lobster giant axon.


1960 ◽  
Vol 43 (5) ◽  
pp. 961-970 ◽  
Author(s):  
John W. Moore ◽  
Kenneth S. Cole

Blood oxygenation and circulation were maintained in Loligo pealii for several hours by a strong flow of sea water over both gills on the open, flat mantle. Potentials were measured with a 3 M KCl-filled glass microelectrode penetrating the giant axon membrane. An hour or more after the mantle was opened, the potentials were similar to those observed in excised axons and in preparations without circulation; spike height 100 mv.; undershoot 12 mv., decaying at 6 v./sec.; resting potential 63 mv. However, the earliest (20 minute) resting potentials were up to 70 mv. and 73 mv. Occasional initial action potential measurements (40 to 50 minute) showed a decay of the undershoot that was less than one-tenth the rate observed later. This suggests that in even better preparations there would be no decay, thereby increasing the resting potential and spike height by 12 mv. With the calculated liquid junction potential of 4 mv. the absolute resting potential in the "normal" axon in vivo is estimated to be about 77 mv., which is close to the Nernst potential for the potassium ratio between squid blood and axoplasm. The differences between such a normal axon and the usual isolated axon can be accounted for by a negligible leakage conductance in the normal axon.


In this part of the discussion we shall attempt to describe the way in which electrical signals are propagated along the giant nerve fibres of squids and cuttlefish. These fibres consist of cylinders of protoplasm, 0.2 to 0.6 mm in diameter, and ire bounded by a thin membrane which acts as a barrier to ionic movement. The protoplasm, or axoplasm as it is commonly called, is an aqueous gel which is a reasonably good conductor of electricity. It contains a high concentration of K + and a low concentration of Na + and Cl - , this situation being the reverse of that in the animal’s blood or sea water. Axons which are left in sea water slowly lose potassium and gain sodium. This process takes about 24 hours and is roughly 80 000 times slower than the diffusion of ions out of a cylinder of gelatin of the same size. The interchange of sodium and potassium is very greatly accelerated by stimulating the fibres. Experiments with tracers, such as those made by Keynes & Lewis (1951) or Rothenberg (1950), allow the interchange to be measured quantitatively, and there is general agreement that the impulse is associated with an entry of 3 to 4 µ µ mol of Na + through 1 cm 2 of membrane and an exit of a corresponding quantity of K + . These quantities are very small compared with the total number of ions inside the fibre. In the giant axon of the squid the quantity of potassium lost in each impulse corresponds to only about 1 millionth if the total internal potassium. One would therefore expect that a giant fibre should be able to carry a great many impulses without recharging its batteries by metabolism. On the other hand, a very small fibre such as a dendrite in the central nervous system should be much more dependent on metabolism since the ratio of surface to volume may be nearly 1000 times greater.


1941 ◽  
Vol 24 (6) ◽  
pp. 771-788 ◽  
Author(s):  
Kenneth S. Cole ◽  
Richard F. Baker

Longitudinal alternating current impedance measurements have been made on the squid giant axon over the frequency range from 30 cycles per second to 200 kc. per second. Large sea water electrodes were used and the inter-electrode length was immersed in oil. The impedance at high frequency was approximately as predicted theoretically on the basis of the poorly conducting dielectric characteristics of the membrane previously determined. For the large majority of the axons, the impedance reached a maximum at a low frequency and the reactance then vanished at a frequency between 150 and 300 cycles per second. Below this frequency, the reactance was inductive, reaching a maximum and then approaching zero as the frequency was decreased. The inductive reactance is a property of the axon and requires that it contain an inductive structure. The variation of the impedance with interpolar distance indicates that the inductance is in the membrane. The impedance characteristics of the membrane as calculated from the measured longitudinal impedance of the axon may be expressed by an equivalent membrane circuit containing inductance, capacity, and resistance. For a square centimeter of membrane the capacity of 1 µf with dielectric loss is shunted by the series combination of a resistance of 400 ohms and an inductance of one-fifth henry.


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.


1978 ◽  
Vol 76 (1) ◽  
pp. 221-235
Author(s):  
J. A. BENSON ◽  
J. E. TREHERNE

The giant axons of this extreme osmoconformer were adapted, in vitro, to progressive hyposmotic dilution of the bathing medium (from 1024 m-Osmol to concentrations as low as 76.8 m-Osmol). Hyposmotic adaptation is associated with reductions in the intracellular concentrations of both sodium and potassium ions. These reductions do not appear to result from appreciable axonal swelling. The different electrical responses to isosmotic and hyposmotic dilution suggest that reduction in [Na+]1 results from ouabain-dependent sodium extrusion, in response to ionic dilution, and that reduction in [K+]1 is induced by a combination of ionic and osmotic dilution. The reduced level of intracellular potassium achieved during hyposmotic adaptation represents a balance between the necessity to contribute to osmotic equilibration and to maintain a potassium gradient across the axon membrane sufficient to produce appreciable axonal hyperpolarization during dilution of the bathing medium. This hyperpolarization tends to maintain the amplitude of the action potential, by compensating for reduction in overshoot (with decline in ENa), and by reducing sodium inactivation. This, together with the reduction in [Na+]1, enables overshooting action potentials of relatively large amplitude and rapid rise time to be maintained during more than tenfold dilution of the ionic and osmotic concentration of the bathing medium.


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