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  • 1
    ISSN: 1432-2013
    Keywords: Concentration-jump technique ; Patch-clamp technique ; Single smooth muscle cell
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A new concentration-jump technique was devised for the rapid application of drugs to single, isolated cells attached to the base of the experimental chamber while recording from them with patch-clamp technique. Cells were placed in a micro-drop (less than 0.1 μl) in a small inner bath which was separated from an outer bath by a ring of “Sylgard” polymer. Stable whole-cell recordings were made in the micro-drop and rapid solution exchange took place when a much larger volume of test solution from the outer bath was flooded over the Sylgard ring and mixed with the micro-drop. Complete equilibration occurred within less than 10 ms.
    Type of Medium: Electronic Resource
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  • 2
    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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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 435 (1998), S. 564-569 
    ISSN: 1432-2013
    Keywords: Key words Patch-clamp ; Ion channel ; Smooth muscle ; Arteriole ; Brain
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The aim of this project was to develop a method to enable routine application of all patch-clamp configurations to smooth muscle cells while they remain embedded in blood vessels. Small blood vessels were isolated from rabbit brain using an enzymatic and mechanical procedure. Vessels were identified under a microscope and the majority were small arterioles with a mean external diameter, in Ca2+-containing (1.5 mM) solution, of 29 μm and variable lengths of 100 μm or more. Arterioles excluded trypan blue, constricted in response to 60 mM K+ and dilated in response to levcromakalim. Patch-clamp gigaOhm seals were made regularly on smooth muscle cells embedded in arterioles. The membrane potential recorded using amphotericin-B-containing patch pipettes averaged –72 mV. Short arteriolar segments could be voltage-clamped. Injection of depolarising current or bath application of 10 mM Ba2+ induced constriction of the entire arteriolar segment. Cell-attached patch, inside-out patch and outside-out patch recordings were made readily and K+ channel unitary currents were studied. The method is readily applied and has several advantages over previous methods for the study of ion channels in smooth muscle cells. Notably, avoidance of single-cell isolation means that enzymatic treatment is minimised and cells can be studied within their normal environment of the blood vessel wall.
    Type of Medium: Electronic Resource
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