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Oxantel-activated single channel currents in the muscle membrane of Ascaris suum

Published online by Cambridge University Press:  06 April 2009

V. M. E. Dale
Affiliation:
Department of Preclinical Veterinary Sciences, R. (D.) S. V. S. Summerhall, University of Edinburgh, Edinburgh EH9 1QH
R. J. Martin
Affiliation:
Department of Preclinical Veterinary Sciences, R. (D.) S. V. S. Summerhall, University of Edinburgh, Edinburgh EH9 1QH

Summary

The patch clamp technique was used to investigate the action of the anthelmintic drug, oxantel, on nicotinic acetylcholine receptor (nAChR) currents recorded from vesicles of the somatic muscle cells of the nematode parasite Ascaris suum. The amplitudes of the currents were analysed at different membrane potentials to determine the single channel conductance. Also the open and closed durations were measured to determine the kinetic properties of the activated channel. Oxantel activated single nAChR currents throughout a concentration range 10–100 μM, these currents were not observed with oxantel-free pipette solutions. The mean open time of the activated channels at a membrane potential of –75 mV and a concentration of 10 μM was 1·34 ms. At higher concentrations the open times were shorter and voltage sensitive, decreasing in duration on hyperpolarization, thus suggesting open channel block. The kinetics were analysed using a simple channel block model. The forward block rate, K + B, increased with increasing oxantel concentration but showed little increase as the membrane was hyperpolarized. K + B was 2·41×107 M−1s−1 – 50 mV and 2·64 × 107 M−1s−1 at – 100mV. The unblocking rate constant, K – B, did exhibit voltage sensitivity being 443·6 s−1 at – 50 mV and 86·8 s−1 at –100 mV. Thus the blocking dissociation constant KB (= K – B/K + B) was 18·5 μM at –50 mV and 3·3 μM at –100 mV. The simple channel block scheme was found to be insufficient to explain fully the observations made; reasons for this are discussed.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1995

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References

REFERENCES

Adams, P. R. (1976). Drug blockade of open end-plate channels. Journal of Physiology 260, 531–52.CrossRefGoogle ScholarPubMed
Adams, D. J., Dwyer, T. M. & Hille, B. (1980). The permeability of endplate channels to monovalent and divalent metal cations. Journal of General Physiology 75, 493510.CrossRefGoogle ScholarPubMed
Cachelin, A. B. & Colquhoun, D. (1989). Desensitization of the acetylcholine receptor of frog end plates measured in a vaseline gap voltage clamp. Journal of Physiology 415, 159–88.CrossRefGoogle Scholar
Colquhoun, D. & Hawkes, A. G. (1982). On the stochastic properties of bursts of single ion channel openings and clusters of bursts. Philosophical Transactions of the Royal Society B300, 159.Google ScholarPubMed
Colquhoun, D., Holden-Dye, L. & Walker, R. J. (1991). The pharmacology of cholinoceptors on the somatic muscle of the parasitic nematode Ascaris suum. Journal of Experimental Biology 158, 509–30.CrossRefGoogle ScholarPubMed
Colquhoun, D. & Sakmann, B. (1981). Fluctuations in the microsecond time range of current through single acetylcholine receptor ion channels. Nature, London 294, 464–6.CrossRefGoogle ScholarPubMed
Colquhoun, D. & Sakmann, B. (1985). Fast events in single channel currents activated by acetylcholine and its analogues at the frog muscle end-plate. Journal of Physiology 369, 501–57.CrossRefGoogle ScholarPubMed
Colquhoun, D. & Sigworth, F. w. (1983). Fitting and single channel analysis of single channel records. In Single Channel Recordings (ed. Sakmann, B. & Nehere, E.), pp. 191264. New York: Plenum Press.CrossRefGoogle Scholar
Hamill, O. P., Marty, A., Neher, E., Sakmann, B. & Sigworth, F. J. (1981). Improved patch clamp techniques for high resolution recording from cells and cell-free membrane patches. Pflügers Archives European Journal of Physiology 391, 85100.CrossRefGoogle ScholarPubMed
Harrow, I. D. & Gration, K. A. F. (1985). Mode of action of the anthelmintics morantel, pyrantel and levamisole on the muscle cell membrane of the nematode Ascaris suum. Pesticide Science 16, 662–72.CrossRefGoogle Scholar
Kuo, C. & Hess, P. (1993). Ion permeation through the L-type calcium channel in rat phaeochromocytoma cells: two sets of ion binding sites in the pore. Journal of Physiology 466, 629–55.CrossRefGoogle ScholarPubMed
Lim, J. K. (1978). Anthelmintic effect of oxantel and oxantel-pyrantel in intestinal nematode infections. Drugs 15 (Suppl 1), 99103.CrossRefGoogle ScholarPubMed
Lindstrom, J., Merlie, J. & Yogeeswaran, G. (1979). Biochemical properties of acetylcholine receptor subunits from Torpedo californica. Biochemistry 18, 4465–70.CrossRefGoogle ScholarPubMed
Martin, R. J. (1985). γ-aminobutyric acid and piperazine activated single channel currents from Ascaris suum body muscle. British Journal of Pharmacology 84, 445–61.CrossRefGoogle ScholarPubMed
Martin, R. J. & Pennington, A. J. (1990). A patch clamp study of acetylcholine activated ion channels in Ascaris suum muscle. Journal of Experimental Biology 154, 201–21.Google Scholar
McFarland, J. W. & Howes, H. L. (1972). Novel anthelmintic agents. 6. Pyrantel analogs with activity against whipworm. Journal of Medicinal Chemistry 15, 365–8.CrossRefGoogle ScholarPubMed
Ogden, D. C. & Colquhoun, D. (1985). Ion channel block by acetylcholine, carbachol and suberyldicholine at the frog neuromuscular junction. Proceedings of the Royal Society of London, B225, 329–55.Google ScholarPubMed
Pennington, A. J. & Martin, R. J. (1990). A patch clamp study of acetylcholine-activated ion channels in Ascaris suum muscle. Journal of Experimental Biology 154, 201–21.CrossRefGoogle ScholarPubMed
Robertson, S. J. & Martin, R. J. (1992). Levamisole-activated single channel currents from muscle of the nematode parasite Ascaris suum. British Journal of Pharmacology 108, 170–8.CrossRefGoogle Scholar
Thorn, P. & Martin, R. J. (1987). A high conductance Ca-dependant chloride channel in Ascaris suum muscle. British Journal of Pharmacology 72, 3149.Google Scholar