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J Physiol. 1978 Jul;280:169-91.

A potential- and time-dependent blockade of inward rectification in frog skeletal muscle fibres by barium and strontium ions.

Abstract

1. A three-electrode voltage clamp method was used to investigate the effects of Ba and Sr ions on the inwardly rectifying K conductance of resting frog sartorius muscle fibres. 2. When Ba2+ (0.01-5 mM) was added to the control (115 mM-K+) solution the inward currents recorded during hyperpolarizing voltage steps turned off exponentially with time as the blockade by Ba2+ developed. Outward currents showed no time-dependence. 3. Ba2+ ions reduced both the instantaneous and the steady-state values of currents recorded on hyperpolarization. The blockade was potential-dependent, steady-state currents being increasingly reduced with increasing hyperpolarization. 4. The concentration-effect relation for the blockade of instantaneous currents by Ba2+ could be fitted assuming 1:1 binding of Ba2+ to a receptor, with the block being proportional to the number of Ba2+-filled receptors. The apparent dissociation constant at the holding potential (-5 mV) was 0.65 mM. Concentration-effect relations were shifted along the concentration axis to lower concentrations by hyperpolarization. The apparent dissociation constant was reduced e-fold for a 16.8 mV change in potential. 5. Increasing the [Ba]o increased the rate of onset of the blockade at a given potential. 6. The rate of onset of the blockade had a high temperature dependence (Q10 = 3.15 +/- 0.08). 7. When [K]o was doubled to 230 mM, under conditions where [K]i was also doubled, [Ba]o had to be raised approximately fourfold to produce the same degree and rate of onset of blockade. Similarly, when [K]o was decreased, the degree and rate of onset of blockade were increased for a given [Ba]o. 8. The blockade could be readily removed by removal of Ba2+ from the bathing solution. In addition the blockade which develops on hyperpolarization is removed exponentially on return to the holding potential. 9. The blockade which exists at the holding potential may be removed by a depolarizing prepulse. 10. Sr causes a similar potential-dependent blockade to that by Ba2+, but is around 400 times less effective. 11. The results have been fitted with a model assuming that the permeability mechanism is an aqueous pore with a site which binds one Ba2+ ion or two K+ ions. The site must have affinity for Ba2+ and a low affinity for K+.

PMID:
308537
PMCID:
PMC1282654
DOI:
10.1113/jphysiol.1978.sp012379
[Indexed for MEDLINE]
Free PMC Article

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