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Electrophysiological and pharmacological characterization of K+-currents in muscle fibres isolated from the ventral sucker of Fasciola hepatica

Published online by Cambridge University Press:  18 November 2004

D. KUMAR ,
C. WHITE ,
I. FAIRWEATHER  and
J. G. McGEOWN
D. KUMAR
Affiliation:
Parasite Proteomics and Therapeutics Research Group, School of Biology and Biochemistry, The Queen's University of Belfast, Belfast BT9 7BL, Northern Ireland
C. WHITE
Affiliation:
Smooth Muscle Research Group, Department of Physiology, The Queen's University of Belfast, Belfast BT9 7BL, Northern Ireland
I. FAIRWEATHER
Affiliation:
Parasite Proteomics and Therapeutics Research Group, School of Biology and Biochemistry, The Queen's University of Belfast, Belfast BT9 7BL, Northern Ireland
J. G. McGEOWN
Affiliation:
Smooth Muscle Research Group, Department of Physiology, The Queen's University of Belfast, Belfast BT9 7BL, Northern Ireland
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Abstract

Fibres isolated from the ventral sucker of Fasciola hepatica were identified as muscle on the basis of their contractility, and their actin and myosin staining. They were voltage-clamped at a holding potential of −40 mV and depolarization-activated outward currents were characterized both electrophysiologically and pharmacologically. Activation was well fitted by a Boltzmann equation with a half-maximal potential of +9 mV and a slope factor of −14·3 mV, and the kinetics of activation and deactivation were voltage-sensitive. Tail current analysis showed that the reversal potential was shifted by +46±3 mV when EK was increased by 52 mV, confirming that this was a K+-current with electrophysiological characteristics similar to delayed rectifier and Ca2+-activated K+-currents in other tissues. The peak current at +60 mV was inhibited by 76±6% by tetrapentylammonium chloride (1 mM) and by 84±7% by Ba2+ (3 mM), but was completely resistant to block by tetraethylammonium (30 mM), 3,4-diaminopyridine (100 μM) and 4-aminopyridine (10 mM). Penitrem A, a blocker of high-conductance Ca2+-activated K+-channels reduced the current at +60 mV by 23±5%. When the effects of Ca2+-channel blocking agents were tested, the peak outward current at +60 mV was reduced by 71±7% by verapamil (30 μM) and by 59±4% by nimodipine (30 μM). Superfusion with BAPTA-AM (50 μM), which is hydrolysed intracellularly to release the Ca2+-buffer BAPTA, also decreased the current by 44±16%. We conclude that voltage-and Ca2+-sensitive K+-channels are expressed in this tissue, but that their pharmacology differs considerably from equivalent channels in other phyla.

Keywords

Fasciola hepaticamuscleactinmyosinimmunocytochemistryscanning electron microscopyvoltage-clampion channels
Type
Research Article
Information
Parasitology , Volume 129 , Issue 6 , December 2004 , pp. 779 - 793
Copyright
© 2004 Cambridge University Press

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References

REFERENCES

AMBROSIO, J., CRUZ-RIVERA, M., ALLAN, J., MORAN, E., ERSFIELD, K. & FLISSER, A. ( 1997). Identification and partial characterization of a myosin-like protein from cysticerci and adults of Taenia solium using a monoclonal antibody. Parasitology 114, 545–553.Google Scholar
ARMSTRONG, C. M. & GILLY, W. F. ( 1992). Access resistance and space clamp problems associated with whole cell patch clamping. Methods in Enzymology 207, 100–122.CrossRefGoogle Scholar
BENNETT, J. L. & KOHLER, P. ( 1987). Fasciola hepatica: action in vitro of triclabendazole on immature and adult stages. Experimental Parasitology 63, 49–57.CrossRefGoogle Scholar
BLAIR, K. L. & ANDERSON, P. A. V. ( 1993). Properties of voltage-activated ionic currents in cells from the brains of the triclad flatworm Bdelloura candida. Journal of Experimental Biology 185, 267–286.Google Scholar
BLAIR, K. L. & ANDERSON, P. A. V. ( 1994). Physiological and pharmacological properties of muscle cells isolated from the flatworm Bdelloura candida (Tricladida). Parasitology 109, 325–335.CrossRefGoogle Scholar
BLAIR, K. L. & ANDERSON, P. A. V. ( 1996). Physiology and pharmacology of turbellarian neuromuscular systems. Parasitology 113, S73–S82.CrossRefGoogle Scholar
BLAIR, K. L., DAY, T. A., LEWIS, M. C., BENNETT, J. L. & PAX, R. A. ( 1991). Studies on muscle cells isolated from Schistosoma mansoni: a Ca2+-dependent K+ channel. Parasitology 102, 251–258.CrossRefGoogle Scholar
BRICKER, C. S., PAX, R. A. & BENNETT, J. L. ( 1982). Microelectrode studies of the tegument and sub-tegumental compartments of male Schistosoma mansoni: anatomical location of sources of electrical potentials. Parasitology 85, 149–161.CrossRefGoogle Scholar
COBBETT, P. & DAY, D. A. ( 2003). Functional voltage-gated Ca2+ channels in muscle fibers of the platyhelminth Dugesia tigrina. Comparative Biochemistry and Physiology A Molecular and Integrative Physiology 134, 593–605.CrossRefGoogle Scholar
COONS, A. H., LEDUC, E. H. & CONNOLLY, J. M. ( 1955). Studies on antibody production. I. A method for the histochemical demonstration of specific antibody and its application to a study of the hyper-immune rabbit. Journal of Experimental Medicine 102, 49–60.Google Scholar
DAY, T. A., KIM, E., BENNETT, J. L. & PAX, R. A. ( 1995). Analysis of the kinetics and voltage-dependency of transient and delayed K+ currents in muscle fibres isolated from the flatworm Schistsoma mansoni. Comparative Biochemistry and Physiology 111A, 79–87.CrossRefGoogle Scholar
DAY, T. A., ORR, N., BENNETT, J. L. & PAX, R. A. ( 1993). Voltage gated currents in muscle cells of Schistosoma mansoni. Parasitology 106, 471–477.CrossRefGoogle Scholar
EDWARDS, G. & WESTON, A. H. ( 1995). The role of potassium channels in excitable cells. Diabetes Research and Clinical Practice 28, S57–S66.CrossRefGoogle Scholar
ETTER, E. F., MINTA, A., POENIE, M. & FAY, F. S. ( 1996). Near-membrane [Ca2+] transients resolved using the Ca2+ indicator FFP18. Proceedings of the National Academy of Sciences, USA 93, 5368–5373.CrossRefGoogle Scholar
FAIRWEATHER, I. & BORAY, J. C. ( 1999). Fasciolicides: efficacy, actions, resistance and its management. Veterinary Journal 158, 81–112.CrossRefGoogle Scholar
HARPER, A. A., CATACUZZENO, L., TREQUATTRINI, C., PETRIS, A. & FRANCIOLINI, F. ( 2001). Verapamil block of large-conductance Ca-activated K channels in rat aortic myocytes. Journal of Membrane Biology 15, 103–111.CrossRefGoogle Scholar
HATANO, N., OHYA, S., MURAKI, K., GILES, W. & IMAIZUMI, Y. ( 2003). Dihydropyridine Ca2+ channel antagonists and agonists block Kv4.2, Kv4.3 and Kv1.4 K+ channels expressed in HEK293 cells. British Journal of Pharmacology 139, 533–544.Google Scholar
HILLE, B. ( 1992). Ionic Channels of Excitable Membrane (2nd Edn), pp. 130–133. Sinauer Associates Inc., Sunderland, Massachusetts.
HORN, R. & MARTY, A. ( 1988). Muscarinic activation of ionic currents measured by a new whole-cell recording method. Journal of General Physiology 92, 145–159.CrossRefGoogle Scholar
JANG, J. H., KIM, S. D., PARK, J. B., HONG, S. J. & RYU, P. D. ( 2003). Ion channels of Fasciola hepatica incorporated into planar lipid bilayers. Parasitology 128, 83–89.Google Scholar
KEENAN, L. & KOOPOWITZ, H. ( 1984). Ionic bases of action potentials in identified flatworm neurones. Journal of Comparative Physiology A 155, 197–208.CrossRefGoogle Scholar
KEENAN, L. & KOOPOWITZ, H. ( 1981). Tetrodotoxin-sensitive action potentials from the brain of the polyclad flatworm Notoplana acticola. Journal of Experimental Zoology 215, 209–213.CrossRefGoogle Scholar
KIM, H. S., KAM, K. Y., RYU, P. D., HONG, S. J., JEON, J. S., JEON, B. H., KIM, K. J. & PARK, J. B. ( 2002). A gadolinium and pH-sensitive hyperpolarization-activated cation current in acutely isolated single neurones from Fasciola hepatica. Parasitology 125, 423–430.CrossRefGoogle Scholar
KNAUS, H.-G., McMANUS, O. B., LEE, S. H., SCHMALHOFER, W. A., GARCIA-CALVO, M., HELMS, L. M. H., SANCHEZ, M., GIANGIACOMO, K., REUBEN, J. P., SMITH, A. B. IIIRD, KACZOROWSKI, G. J. & GARCIA, M. L. ( 1994). Tremorgenic indole alkaloids potently inhibit smooth muscle high-conductance calcium-activated potassium channels. Biochemistry 33, 5819–5828.CrossRefGoogle Scholar
KUMAR, D., McGEOWN, J. G., REYNOSO-DUCOING, O., AMBROSIO, J. R. & FAIRWEATHER, I. ( 2003). Observations on the musculature and isolated muscle fibres of the liver fluke, Fasciola hepatica. Parasitology 127, 457–473.CrossRefGoogle Scholar
LARGE, W. A. & WANG, Q. ( 1996). Characteristics and physiological role of the Ca2+-activated Cl− conductance in smooth muscle. American Journal of Physiology 271, C435–C454.CrossRefGoogle Scholar
LINCKS, J., BOYAN, B. D., BLANCHARD, C. R., LOHMANN, C. H., LIU, Y., COCHRAN, D. L., DEAN, D. D. & SCHWARTZ, Z. ( 1998). Response of MG63 osteoblast-like cells to titanium and titanium alloy is dependent on surface roughness and composition. Biomaterials 19, 2219–2232.CrossRefGoogle Scholar
LOIRAND, G., PACAUD, P., BARON, A., MIRONNEAU, C. & MIRONNEAU, J. ( 1991). Large conductance calcium-activated non-selective cation channel in smooth muscle cells isolated from rat portal vein. Journal of Physiology 437, 461–475.CrossRefGoogle Scholar
MARTIN, R. J. ( 1997). Modes of action of anthelminthic drugs. Veterinary Journal 154, 11–34.CrossRefGoogle Scholar
MURAKUMO, M., USHIKI, T., ABE, K., MATSUMARA, K., SHINNO, Y. & KOYANAGI, T. ( 1995). Three-dimensional arrangement of collagen and elastin fibers in the human urinary bladder: a scanning electron microscopic study. Journal of Urology 154, 251–256.CrossRefGoogle Scholar
NEHER, E. ( 1992). Correction for liquid junction potentials in patch clamp experiments. Methods in Enzymology 207, 123–131.CrossRefGoogle Scholar
NELSON, M. T., PATLAK, J. B., WORLEY, J. F. & STANDEN, N. B. ( 1990). Calcium channels, potassium channels, and voltage dependence of arterial smooth muscle tone. American Journal of Physiology 259, C3–C18.CrossRefGoogle Scholar
SAKMANN, B. & NEHER, E. ( 1984). Patch clamp techniques for studying ionic channels in excitable membranes. Annual Review of Physiology 46, 455–472.CrossRefGoogle Scholar
SOLON, M. & KOOPOWITZ, H. ( 1982). Multimodal interneurones in the polyclad flatworm Alloeoplana californica. Journal of Comparative Physiology A 147, 171–178.CrossRefGoogle Scholar
THOMSON, S. H. ( 1977). Three pharmacologically distinct potassium channels in molluscan neurones. Journal of Physiology 265, 465–488.CrossRefGoogle Scholar
URBANO, F. J. & BUNO, W. ( 1998). BAPTA-AM blocks both voltage gated and Ca2+-activated K+ currents in cultured bovine chromaffin cells. Neuroreport 9, 3403–3407.CrossRefGoogle Scholar
VOLK, K. A., MATSUDA, J. J. & SHIBATA, E. F. ( 1991). A voltage-dependent potassium current in rabbit coronary artery smooth muscle cells. Journal of Physiology 439, 751–768.CrossRefGoogle Scholar
ZHABYEYEV, P., MISSAN, S., JONES, S. E. & McDONALD, T. F. ( 2000). Low-affinity block of cardiac K+ currents by nifedipine. European Journal of Pharmacology 401, 137–143.CrossRefGoogle Scholar
ZHANG, S., ZHOU, Z., GONG, Q., MAKIELSKI, J. C. & JANUARY, C. T. ( 1999). Mechanism of block and identification of the verapamil binding domain to HERG potassium channels. Circulation Research 84, 989–998.CrossRefGoogle Scholar