Transmembrane Activity Gradients of K and cl in the Muscle Fibres of Osmoconforming Marine Decapod Crustaceans

1978 ◽  
Vol 72 (1) ◽  
pp. 127-140
Author(s):  
ROBERT W. FREEL

1. The resting membrane potentials (Em) and the transmembrane activity gradients for K and Cl were measured in the muscle fibres of osmoconforming (Callianassa and Cancer) and weakly osmoregulating (Pachygrapsus) marine crustaceans acclimated to various osmotic conditions. 2. The muscle membranes of sea water acclimated crabs behave as good K electrodes. However, a slight contribution of Na to the resting potential was demonstrated in all species. The ratio PNa/PK was about 0.01. Equilibrium potentials (measured with ion-selective microelectrodes) for Cl were equal to Em, while EK was always more negative than Em as a result of the slight Na contribution. 3. Acclimation to dilute or concentrated sea water had little effect on the K electrode properties or Na permeabilities of these fibres. The muscle fibres were depolarized in crabs acclimated to concentrated sea water and were hyperpolarized in crabs adapted to dilute sea water. These changes resulted solely from alterations in (aK)i/ (aK)O which were in turn brought about by changes in cellular and haemolymph hydration. 4. Since the Na contribution to Em was so small in all conditions, it was concluded that the distributions of K and Cl are best considered in terms of Donnan equilibria, and that the cellular K and Cl adjustments observed during salinity adaptation reflect the passive re-establishment of new equilibrium distributions for these ions.

1991 ◽  
Vol 155 (1) ◽  
pp. 305-321
Author(s):  
D. A. DORSETT ◽  
C. G. EVANS

Dissociated unstriated muscle fibres from the buccal mass retractor muscles of the mollusc Philine aperta were studied using a two-electrode voltage-clamp. The mean resting potential of the fibres was −76.3±0.44mV (N=30), and the membrane resistance was 42.2±3MΩ. The space constant of the fibres was 2.03+0.33mm (N=5). Three outward potassium currents were resolved in response to a depolarising step to zero from resting potential. (1) An early transient current, voltageactivated and blocked by 2 mmoll−1 4-aminopyridine (4-AP). This resembled the A-current described in molluscan neurones and some arthropod muscle fibres. (2) A calcium-dependent late transient current, with slower kinetics, which was suppressed by 50 mmoll−1 tetraethylammonium chloride (TEA-Cl), zero-calcium saline, 1 mmol−1 Cd2+ and 1 μmoll−1 verapamil. (3) A delayed voltage-activated current, blocked by 50 mmoll−1 TEA-Cl and with kinetics associated with the delayed rectifier current IK. An inwardly directed current, blocked by zero-calcium salines, Cd2+ and verapamil, was considered to be a calcium current whose activation closely matched that of the Ca2+-dependent potassium current. A blockade of either the A-current, or exposure to low-calcium artificial sea water, or a combination of both, promoted the development of oscillations and regenerative spikes in the muscle fibre following depolarization


1958 ◽  
Vol 41 (3) ◽  
pp. 529-542 ◽  
Author(s):  
John C. Dalton

The effects of varying external concentrations of normally occurring cations on membrane potentials in the lobster giant axon have been studied and compared with data presently available from the squid giant axon. A decrease in the external concentration of sodium ions causes a reversible reduction in the amplitude of the action potential and its rate of rise. No effect on the resting potential was detected. The changes are of the same order of magnitude, but greater than would be predicted for an ideal sodium electrode. Increase in external potassium causes a decrease in resting potential, and a decrease in potassium causes an increase in potential. The data so obtained are similar to those which have been reported for the squid giant axon, and cannot be exactly fitted to the Goldman constant field equation. Lowering external calcium below 25 mM causes a reduction in resting and action potentials, and the occasional occurrence of repetitive activity. The decrease in action potential is not solely attributable to a decrease in resting potential. Increase of external calcium from 25 to 50 mM causes no change in transmembrane potentials. Variations of external magnesium concentration between zero and 50 mM had no measurable effect on membrane potentials. These studies on membrane potentials do not indicate a clear choice between the use of sea water and Cole's perfusion solution as the better external medium for studies on lobster nerve.


1978 ◽  
Vol 72 (1) ◽  
pp. 107-126
Author(s):  
ROBERT W. FREEL

1. The patterns of myoplasmic water and solute regulation were examined in several species of marine decapod crustaceans acclimated to various salinities. Three osmoconformers (Callianassa cahformemsis, Cancer antennarius, Emerita analoga) and one weak osmoregulator (Pachygrapsus crassipes) were studied. 2. Although haemolymph water and solute activities varied with adaptational salinity in these species, regulation of cell water content was apparent in all but 25 % s.w. acclimated Pachygrapsus. Cell water regulation was correlated with the regulation of ninhydrin-positive solutes. An estimate of the extent of the regulation was achieved by comparing the observed cell hydration changes to that predicted for an ideal osmometer and to that of a perfect volume regulator. It was concluded that the extent of cell volume regulation was variable and incomplete, particularly in dilute salinities. 3. Net losses of myoplasmic sodium, chloride, and ninhydrin-positive nitrogen were observed in animals acclimated to dilute sea water, while net gains of these solutes were noted in crabs acclimated to concentrated sea water. In contrast, the amounts of cell potassium and calcium plus magnesium were relatively constant fractions of the cell dry weight at any salinity. 4. A simple mechanism, br.sed upon Donnan considerations, is sufficient to account for the constancy of myoplasmic potassium and the variability of cell chloride with changing extracellular osmolarity.


1992 ◽  
Vol 68 (4) ◽  
pp. 985-1000 ◽  
Author(s):  
H. Sontheimer ◽  
J. A. Black ◽  
B. R. Ransom ◽  
S. G. Waxman

1. Na+ and K+ channel expression was studied in cultured astrocytes derived from P--0 rat spinal cord using whole cell patch-clamp recording techniques. Two subtypes of astrocytes, pancake and stellate, were differentiated morphologically. Both astrocyte types showed Na+ channels and up to three forms of K+ channels at certain stages of in vitro development. 2. Both astrocyte types showed pronounced K+ currents immediately after plating. Stellate but not pancake astrocytes additionally showed tetrodotoxin (TTX)-sensitive inward Na+ currents, which displayed properties similar to neuronal Na+ currents. 3. Within 4-5 days in vitro (DIV), pancake astrocytes lost K(+)-current expression almost completely, but acquired Na+ currents in high densities (estimated channel density approximately 2-8 channels/microns2). Na+ channel expression in these astrocytes is approximately 10- to 100-fold higher than previously reported for glial cells. Concomitant with the loss of K+ channels, pancake astrocytes showed significantly depolarized membrane potentials (-28.1 +/- 15.4 mV, mean +/- SD), compared with stellate astrocytes (-62.5 +/- 11.9 mV, mean +/- SD). 4. Pancake astrocytes were capable of generating action-potential (AP)-like responses under current clamp, when clamp potential was more negative than resting potential. Both depolarizing and hyperpolarizing current injections elicited overshooting responses, provided that cells were current clamped to membrane potentials more negative than -70 mV. Anode-break spikes were evoked by large hyperpolarizations (less than -150 mV). AP-like responses in these hyperpolarized astrocytes showed a time course similar to neuronal APs under conditions of low K+ conductance. 5. In stellate astrocytes, AP-like responses were not observed, because the K+ conductance always exceeded Na+ conductance by at least a factor of 3. Thus stellate spinal cord astrocyte membranes are stabilized close to EK as previously reported for hippocampal astrocytes. 6. It is concluded that spinal cord pancake astrocytes are capable of synthesizing Na+ channels at densities that can, under some conditions, support electrogenesis. In vivo, however, AP-like responses are unlikely to occur because the cells' resting potential is too depolarized to allow current activation. Thus the absence of electrogenesis in astrocytes may be explained by two mechanisms: 1) a low Na-to-K conductance ratio, as in stellate spinal cord astrocytes and in other previously studied astrocyte preparations; or, 2) as described in detail in the companion paper, a mismatch between the h infinity curve and resting potential, which results in Na+ current inactivation in spinal cord pancake astrocytes.


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.


1981 ◽  
Vol 94 (1) ◽  
pp. 251-268
Author(s):  
LUDWIG FISCHER ◽  
ERNST FLOREY

In experiments on the opener muscle of the third walking legs of crayfish (Astacus leptodactylus) it was found that the mechanical tension developed in response to repetitive stimulation of the single motor axon increases over the entire temperature range from 30 down to 0°C. In contrast to this, the tension elicited by depolarizing single muscle fibres decreases with decreasing temperature; the threshold for excitation-contraction coupling is not significantly altered. With decreasing temperature the resting potential decreases (up to 2 mV/°C) but the amplitude and decay time of the e.p.s.p.'s increase. The time constant, λ, of e.p.s.p. decay has a Q10 of less than −2 in the range above 15 °C but reaches a value of −7 between 10 and 0°C. This pattern of temperature dependence is fully accounted for by a parallel change of membrane resistance and its reciprocal, the membrane conductance. The corresponding activation energies computed from λ-values approximate 3 kcal/mol at high temperature and 46 kcal/mol in the low temperature range. The combined effects of a lowered resting potential, an increased amplitude, and especially an increased decay time of e.p.s.p.s result in a drastic enhancement of the depolarization reached during summation of e.p.s.p.s as the temperature is decreased. These effects overcompensate the declining effectiveness of excitation-contraction coupling so that the entire process of neuromuscular transmission becomes more and more effective as the temperature declines. In order to reach the same tension lower frequencies of nerve stimulation are needed at lower temperatures.


1990 ◽  
Vol 63 (1) ◽  
pp. 72-81 ◽  
Author(s):  
A. Williamson ◽  
B. E. Alger

1. In rat hippocampal pyramidal cells in vitro, a brief train of action potentials elicited by direct depolarizing current pulses injected through an intracellular recording electrode is followed by a medium-duration afterhyperpolarization (mAHP) and a longer, slow AHP. We studied the mAHP with the use of current-clamp techniques in the presence of dibutyryl cyclic adenosine 3',5'-monophosphate (cAMP) to block the slow AHP and isolate the mAHP. 2. The mAHP evoked at hyperpolarized membrane potentials was complicated by a potential generated by the anomalous rectifier current, IQ. The mAHP is insensitive to chloride ions (Cl-), whereas it is sensitive to the extracellular potassium concentration ([K+]o). 3. At slightly depolarized levels, the mAHP is partially Ca2+ dependent, being enhanced by increased [Ca2+]o and BAY K 8644 and depressed by decreased [Ca2+]o, nifedipine, and Cd2+. The Ca2(+)-dependent component of the mAHP was also reduced by 100 microM tetraethylammonium (TEA) and charybdotoxin (CTX), suggesting it is mediated by the voltage- and Ca2(+)-dependent K+ current, IC. 4. Most of the Ca2(+)-independent mAHP was blocked by carbachol, implying that IM plays a major role. In a few cells, a small Ca2(+)- and carbachol-insensitive mAHP component was detectable, and this component was blocked by 10 mM TEA, suggesting it was mediated by the delayed rectifier current, IK. The K+ channel antagonist 4-aminopyridine (4-AP, 500 microM) did not reduce the mAHP. 5. We infer that the mAHP is a complex potential due either to IQ or to the combined effects of IM and IC. The contributions of each current depend on the recording conditions, with IC playing a role when the cells are activated from depolarized potentials and IM dominating at the usual resting potential. IQ is principally responsible for the mAHP recorded at hyperpolarized membrane potentials.


1962 ◽  
Vol 45 (6) ◽  
pp. 1195-1216 ◽  
Author(s):  
Fred J. Julian ◽  
John W. Moore ◽  
David E. Goldman

A method similar to the sucrose-gap technique introduced be Stäpfli is described for measuring membrane potential and current in singly lobster giant axons (diameter about 100 micra). The isotonic sucrose solution used to perfuse the gaps raises the external leakage resistance so that the recorded potential is only about 5 per cent less than the actual membrane potential. However, the resting potential of an axon in the sucrose-gap arrangement is increased 20 to 60 mv over that recorded by a conventional micropipette electrode when the entire axon is bathed in sea water. A complete explanation for this effect has not been discovered. The relation between resting potential and external potassium and sodium ion concentrations shows that potassium carries most of the current in a depolarized axon in the sucrose-gap arrangement, but that near the resting potential other ions make significant contributions. Lowering the external chloride concentration decreases the resting potential. Varying the concentration of the sucrose solution has little effect. A study of the impedance changes associated with the action potential shows that the membrane resistance decreases to a minimum at the peak of the spike and returns to near its initial value before repolarization is complete (a normal lobster giant axon action potential does not have an undershoot). Action potentials recorded simultaneously by the sucrose-gap technique and by micropipette electrodes are practically superposable.


1994 ◽  
Vol 71 (2) ◽  
pp. 575-582 ◽  
Author(s):  
E. Puil ◽  
H. Meiri ◽  
Y. Yarom

1. We studied the voltage responses of thalamocortical neurons to a periodic current input of variable frequency, in slices of mediodorsal thalamus (guinea pig). The ratio of the Fourier transform of the voltage response to the Fourier transform of the oscillatory current input was used to calculate the frequency response of the neurons at different resting and imposed membrane potentials. 2. Most neurons displayed a resonant hump in the frequency response curve. A narrow band of low-frequency (2-4 Hz) resonance occurred near the resting level [-66 +/- 8 mV (SD)] and at imposed membrane potentials in a range of -60 to -80 mV. An additional wide band (12-26 Hz) of peak resonant frequencies was observed at depolarized levels. 3. The low-frequency resonance was insensitive to tetrodotoxin (TTX) application in concentrations (0.5-1 microM) that blocked a depolarization activated inward rectifier and Na(+)-dependent action potentials. TTX, however, eliminated the wide-band resonant hump centered at 12-26 Hz that we observed at depolarized membrane potentials. 4. Application of Ni2+ (0.5-1 mM) reversibly blocked all slow spikes and greatly reduced the low-frequency resonant humps, without changing the resting potential. Octanol in concentrations of 50 microM had similar effects. 5. Application of Cs+ (3-5 mM), a blocker of the hyperpolarization activated inward rectifier, produced a 5- to 10-mV depolarization and completely blocked the rectification. Cs+ did not alter the low-frequency resonant hump or its dependence on membrane voltage.(ABSTRACT TRUNCATED AT 250 WORDS)


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