Axonal accessibility and adaptation to osmotic stress in an extreme osmoconformer.
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Biomedical subjects
Publications and source records attributed to J E Treherne.
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Despite the extreme fluctuations in blood concentration experienced by this marine osmoconformer, essentially 'conventional' ionic mechanisms are involved in conduction by the giant axons in isosmotic conditions. The resting axonal membrane approximates to an ideal potassium electrode, with a 58-8 mV slope for decade change in [K+]O above 10 mM. The action potential overshoot shows a 55-8 mV decline with decade reduction in [Na+]O and the action potentials are blocked by tetrodotoxin at around 5 X 10(-7) M. The rising phase and overshoot of the action potential remain constant at potassium concentrations up to the relatively high level of 30 mM found in the blood, indicating an unusual absence of sodium inactivation over a wide range of resting potentials. Relatively rapid, symmetrical movement of potassium ions between the bathing medium and the axon surface is deduced from the potential changes induced by alterations in [K+]O. Outward movement of sodium ions (t0-5 = 33-5 s) occurs at a similar rate to that of potassium, but inward movement of Na+ is relatively slow and complex. It is concluded that the ability of axons to function in dilute media must involve specific adaptations to osmotic and ionic stress.
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In contrast to the depolarization observed in hyperosmotic media, exposure of peripheral nerve to hyposmotic conditions induced pronounced axonal hyperpolarization. It is suggested that this hyperpolarization resulted from increased potassium and chloride permeabilities which could assist axonal volume regulation in hyposmotic conditions. The hyperpolarization was readily reversible, but the spike-generating mechanism suffered irreversible damage at hyposmotic concentrations below 665 m-osmoles. It is suggested that this axonal damage contributes to the lethal effects of hyposmotic stress in this crustacean osmoconformer and, possibly, in some euryhaline osmoregulators.
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Exposure to sodium-deficient (tris) saline caused an appreciable decline in the sodium content of intact connectives in the absence of equivalent reduction in the amplitude of the recorded action potentials. Return of sodium-depleted connectives to normal saline resulted in a rapid recovery of axonal function despite only a partial (less than 70%) recovery in sodium content. Replacement of sodium ions by those of lithium in the bathing medium resulted in a substantial accumulation of this cation. Lithium movements exhibited a marked asymetry, no significant decline in concentration being observed upon return to normal saline. These results are tentatively interpreted in terms of an exchangeable glial sodium fraction and are discussed in relation to extra-axonal sodium regulation.
The apparent activity coefficients for sodium (gamma'Na) in the blood of six insect species have been calculated from measurements made with sodium-selective electrodes and a flame photometer. In every case gamma Na was significantly lower than that for this cation in free solution (gammaNa). In Periplaneta americana gammaNa varied considerably, during a period of 90 days, so that a relatively constant sodium activity (aNa) was maintained in the blood in the face of large variations in the total sodium content measured by flame photometry. Despite the relative constancy of aNa (of around 0.088M) appreciable fluctuations were observed in the sodium and potassium content of nervous connective over a period of 140 days. The values of aNa and aK were used to devise a satisfactory cockroach saline for use in experiments with isolated nerve cords.
An accelerated water-loss was observed in decapitated individuals, no equivalent increase being obtained following severance of the nervous connectives in the neck. Injection of brain and, to a lesser extent, corpus cardiacum extract resulted in a significant reduction in the rate of loss of water from decapitated individuals. The accelerated water-loss observed following decapitation appeared not to result from significant increase in excretory output or loss of water through the spiracles. It is suggested that integumentary transpiration may be affected by a blood-borne factor, or factors, which originate in the brain and corpus cardiacum.
Simultaneous intracellular and sucrose-gap recordings showed, in contrast to previous findings, that the electrical parameters of giant axons were similar to intact and desheathed connectives bathed with the 'extracellular Ringer' of Yamasaki & Narahashi. This implies that the extra-axonal sodium concentration, in situ, is likely to be lower than had been previously supposed. Axonal responses showed that, despite the high blood concentration of 24-2 mM-K+ measured by flame photometry, the effective concentration in the blood was 10-15 mM-K+ which corresponds to the measurements made with potassium-selective electrodes. The activity of the blood potassium ions caused a marked reduction in the amplitude of the action potentials following surgical desheathing or disruption of the blood-brain barrier with hypertonic urea. It is suggested that a regulatory mechanism exists in the central nervous system which counteracts the effects of the high blood potassium level.
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