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  • 1
    ISSN: 1432-2013
    Keywords: concentration-jump technique ; patch-clamp technique ; single smooth muscle cell ; 1,4-dihydropyridines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The actions of nifedipine or BAY K 8644 were studied on barium currents recorded from single, collagenase- and elastase-dispersed, smooth muscle cells from the rabbit ear artery using the whole-cell configuration of the patch-clamp technique. Nifedipine (3μM) caused a reduction in the barium current (IBa) evoked by steps to potentials positive of-10mV. This was characterized by a pronounced ‘initial’ block, an increase in the rate of current decay during the voltage-clamp step, but by no increase in block if pulses were repeated every 600ms. Rapid extracellular application of nifedipine (1μM) during the sustained current component (using a new concentration-jump technique) was found to have no effect on IBa over 4s at +20mV, but after returning to the holding potential (-60mV) for 10s, sustained IBa was subsequently abolished. BAY K 8644 (1μM) increased IBa at all potentials, and on rapid application during the sustained current component markedly potentiated IBa. The results suggest that nifedipine binds with high affinity to the closed, available state of the Ca++ channels but they do not suggest binding to the open or inactivated states. The effect of BAY K 8644 is consistent with high affinity binding to the open or inactivated and to the closed, available states.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: Calcium oscillations ; Muscarinic receptor ; Calcium stores ; G protein ; Heparin ; Ryanodine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In single cells isolated from guinea-pig ileal smooth muscle, held under voltage clamp at −40 mV or −50 mV by patch pipette in the whole-cell recording mode, carbachol (CCh) evoked an oscillatory inward cationic current. The frequency of current oscillations increased with increasing CCh concentration. CCh-evoked current oscillations were followed very closely by oscillations in intracellular free Ca2+ estimated from the Indo-1 signal, and were abolished by inclusion of EGTA in the pipette solution. Ryanodine and heparin, but not nifedipine, blocked the generation of current oscillations. CCh-evoked current oscillations were abolished upon withdrawal of extracellular calcium and restored upon its reintroduction. Inclusion of GTP[γS] in the pipette solution caused the generation of an oscillatory inward current, which was blocked by ryanodine. The present results are consistent with the hypothesis that CChevoked cationic current is gated by activation of a G protein and is steeply dependent on [Ca2+]i, fluctuations in the release of Ca2+ from stores during carbachol's action produce oscillations in [Ca2+]i which cause similar oscillations in the cationic current.
    Type of Medium: Electronic Resource
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