4.4 Article

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

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PARASITOLOGY
卷 129, 期 -, 页码 779-793

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CAMBRIDGE UNIV PRESS
DOI: 10.1017/S0031182004006110

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Fasciola hepatica; muscle; actin; myosin; immunocytochemistry; scanning electron microscopy; voltage-clamp; ion channels

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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 E-K was increased by 52 mV, confirming that this was a K+-current with electrophysiological characteristics similar to delayed rectifier and Ca-2+-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 mum) 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 mum) and by 59 +/- 4% by nimodipine (30 mum). Superfusion with BAPTA-AM (50 muM), 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.

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