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  • 1
    ISSN: 1432-1424
    Keywords: patch clamp ; epithelial ion transport ; quinidine ; divalen cation blockade ; ion channels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Outward rectifying. cation channels were observed in the epithelial cells of the urinary bladder of the toad.Bufo marinus. As studied in isolated cells using the patch-clamp technique, the channel has an average conductance of 24 and 157 pS for pipette potentials between 0 and +60 mV and −60 to −100 mV, respectively, when the major cation in both bath and pipette solutions is K+. The conductance of the cannel decreasen with increasing dehydration energy of the permeant monovalent cation in the oder Rb+=K+〉Na+〉Li+. Reversal potentials near zero under biionic conditions imply that the permeabilities for all four of these cations are smiliar. The channel is sensitive to quinidine sulfate but not to amiloride. It shares several pharmacological and biophysical properties with an outwardly-rectifying, vasopressin-sensitive pical K+ conductive pathway described previously for the toad urinary bladder. We demonstrate, in both single-channel and whole-bladder studies, that the outward rectification is a consequence of interaction of the chanel with extracellular divalent cations, particularly Ca2+, which blocks inward but not outward current. Various divalent cations impart different degrees of outward rectification to the conductive pathway. Concentrations of Mg2+ and Ca2+ required for halfmaximal effect are 3×10−4 and 10−4 m, resopectively. For Co2+ the values are 10−6 m at +50 mV and a 10−4 m at +200 mV. The mechanism of blockade by divalent cations is not established, but does not seem to involve a voltage-dependent interaction in which the blocker penetrates the transmembrane electric field. In the absence of divalent cations in the mucosal solution, the magnitudes of inward current carried by Rb+, K+, Na+ and Li+ through the apical K+ pathway at any transepithelial voltage, are in the same order as in the single-channel studies. We propose that the cation channel observed by us in isolated epithelial cells is the single-channel correlate of the vasopressin-sensitive apical K+ conductive pathway in the toad urinary bladder and is also related to the oxytocin- and divalent cation-sensitive apical condictivity observed in frog skin and urinary bladder.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 92 (1986), S. 217-226 
    ISSN: 1432-1424
    Keywords: toad urinary bladder ; K channels ; ADH ; carbachol ; Li ; quinidine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The conductance of the apical membrane of the toad urinary bladder was studied under voltage-clamp conditions at hyperpolarizing potentials (mucosa negative to serosa). The serosal medium contained high KCl concentrations to reduce the voltage and electrical resistance across the basal-lateral membrane, and the mucosal solution was Na free, or contained amiloride, to eliminate the conductance of the apical Na channels. As the mucosal potential (V m) was made more negative the slope conductance of the epithelium increased, reaching a maximum at conductance of the epithelium increased, reaching a maximum atV m=−100 mV. This rectifying conductance activated with a time constant of 2 msec whenV m was changed abruptly from 0 to −100 mV, and remained elevated for at least 10 min, although some decrease of current was observed. ReturningV m to+100 mV deactivated the conductance within 1 msec. Ion substitution experiments showed that the rectified current was carried mostly by cations moving from cell to mucosa. Measurement of K flux showed that the current could be accounted for by net movement of K across the apical membrane, implying a voltage-dependent conductance to K (G K). Mucosal addition of the K channel blockers TEA and Cs had no effect onG K, while 29mm Ba diminished it slightly. Mucosal Mg (29mm) also reducedG K, while Ca (29mm) stimulated it.G K was blocked by lowering the mucosal pH with an apparent pK1 of 4.5. Quinidine (0.5mm in the serosal bath) reducedG K by 80%.G K was stimulated by ADH (20 mU/ml), 8-Br-cAMP (1mm), carbachol (100 μm), aldosterone (5×10−7 m for 18 hr), intracellular Li and extracellular CO2.
    Type of Medium: Electronic Resource
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