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  • Caged ATP  (1)
  • Calcium current  (1)
  • Carbachol  (1)
  • Concentration-jump technique  (1)
  • 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: Smooth Muscle ; Slow Waves ; Carbachol ; Role of Ions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The membrane potential of the cells of the longitudinal muscle of the guinea-pig ileum was recorded intracellularly with glass microelectrodes. Upon changing from isotonic physiological salt solution to sucrose hypertonic solution the spontaneous electrical activity of the membrane was abolished. Spike discharge, but not slow potential changes, was evoked by depolarizing current. In isotonic or in sucrose hypertonic solution, carbachol or acetylcholine caused spike discharge and produced oscillations of the membrane potential (slow waves) which, in hypertonic solution, were about 20 mV in size and 3 sec in duration. The effects on the response to carbachol of varying the ionic composition were examined in sucrose-hypertonic solution. Slow waves in response to carbachol were rapidly and reversibly abolished in sodium-deficient solution, though electrical stimulation evoked spikes for considerable periods. Slow waves were abolished also in sodium-free solution. In contrast, carbachol evoked slow waves after 20 min in calcium-free solution (in which the membrane depolarized) if the membrane was electrically repolarized. In chloride-deficient solution a small but significant (p〈0.05) increase occurred in the duration of slow waves evoked by carbachol. Carbachol elicited slow waves in potassium-free or in potassium-rich solution. The increases in slow wave size and duration in potassium-free solution fell short of statistical significance (0.1〉p〉0.05). The depolarization produced by carbachol was significantly (p〈0.05) less in sodium-deficient (15 mM) solution but was unaffected by alterations in the external chloride concentrations. In sodium-free solution, carbachol hyperpolarized the membrane. The results support a previous suggestion that the slow waves produced by acetylcholine or carbachol represent an inward sodium current through a slow regenerative ion channel.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2013
    Keywords: Inositol trisphosphate ; Caged InsP 3 ; Caged ATP ; Heparin ; Calcium current
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract In single cells obtained by enzymic treatment of rabbit small-intestinal smooth muscle, and held under voltage clamp by patch pipette in the whole-cell recording mode, release of inositol trisphosphate (InsP 3) from its caged precursor by flash photolysis caused complete inhibition of the voltage-dependent calcium current. No inhibition was seen in control experiments where the cage (2-nitrosoacetophenone) was released by flash photolysis from caged ATP. The inhibition by InsP 3 of the calcium current was prevented if 10 mM EGTA or 2 mg/ml heparin was included in the pipette solution. Heparin is known to block InsP 3 receptors. These results suggest that release of calcium stores by InsP 3 raises Cai and that calcium ions inhibit the calcium current by acting either directly or otherwise on the internal mouth of the calcium channel.
    Type of Medium: Electronic Resource
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