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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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