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  • 1
    ISSN: 1432-2013
    Keywords: Collecting duct ; Principal cell ; Tissue culture ; Aldosterone ; Amiloride ; Sodium transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Whereas collecting duct epithelium in vivo is composed of principal and intercalated cells, we grew a pure principal cell epithelium using a new technique involving tissue culture. These principal cells were derived from collecting duct anlagen of newborn rabbits. We investigated the electrical properties of such epithelia in a newly designed lucite double-chamber with an inner opening of 0.08 cm2. Our observations were: 1) mean transepithelial resistanceR te was 0.83±0.2 kΩcm2 at 37° C and after preincubation in aldosterone; 2) mean transepithelial potential differenceV te was low and variable under standard conditions and at room temperature but increased to −59.5±4.4 mV (sign referring to polarity of apical surface) after preincubation in 10−6 mol/l aldosterone and at 37° C; 3) 10−6 mol/l amiloride added to the apical perfusion fluid largely abolished thisV te while increasingR te by 120%; 4) experiments with 5×10−3 mol/l BaCl2 in the apical perfusion fluid failed to changeR te andV te significantly. This principal cell epithelium therefore has characteristics of a “tight” epithelium with active sodium transport; however, its electrical properties differ from those of the isolated perfused collecting duct segment.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2013
    Keywords: Collecting duct ; Principal cell ; Tissue culture ; Chloride conductivity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The ionic conductive properties were studied of epithelia of collecting duct principal cells which had been grown in primary tissue culture from renal cortex/capsule explants. When pretreated with aldosterone (10−6 mol/l) and bathed on either surface with isotonic HCO 3 − -free Ringer's solution, the transepithelial voltage,V te, varied between −21 and −72 mV (apical surface negative) while the transepithelial resistance,R te, ranged from 0.4 to 1.5 kΩcm2. By 10:1 step-changes in Na+ concentration the apical cell membrane was shown to have a high conductivity for sodium, inhibitable by amiloride, 10−6 mol/l. However, contrary to observations in natural collecting duct under control conditions, amiloride never reversed the polarity ofV te even at 10−4 mol/l. Both the apical and the basolateral cell membranes were conductive for potassium and both conductivities were inhibitable by Ba2+ (5 mmol/l). 10:1 reduction of apical Cl− concentration strongly hyperpolarizedV te with a monophasic time course suggesting the presence of a paracellular shunt conductance for Cl−. In addition there may be a small Cl− conductance present in the apical cell membrane since apical application of the chloride channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPAB) at 10−7 mol/l produced a minute but significant hyperpolarization. On the other hand, 10:1 reduction of basolateral Cl− concentration caused a biphasic change inV te (initial depolarization, followed by repolarization) which indicates the presence of a large Cl− conductance in the basolateral cell membrane. The latter was not inhibitable by 10−7 mol/l NPPAB. Higher concentrations of this and of an other Cl− channel blocker produced non-specific effects. In conclusion, our studies of a pure principal cell epithelium confirm findings described for the intact cortical collecting duct and add new information concerning chloride conductivity and related blocking agents.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2013
    Keywords: Collecting duct principal cell culture ; High conductance K+ channel ; Renal K+ secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Using the patch clamp technique, one type of K+ channel was identified in the apical cell membrane of cultured principal cells of rabbit renal collecting ducts in the cell-attached or excised-patch configuration. The channel was highly selective for K+ over Na+ (typically 30-70-fold) and had a conductance of 180, SD±39 pS (n=6), referred to a situation of 140 mmolar K+-Ringer solution present on either surface of the patch membrane. Channel activity was completely blocked by Ba2+ (5 mmol/l) and partially inhibited by Na+. The latter was evidenced by a deviation from Goldman rectification at high cytoplasm-positive membrane potentials, which was observed when Na+ competed with K+ for channel entrace from the cytoplasmic surface. Channel open probability depended strongly on membrane voltage and cytoplasmic Ca2+ concentration. Open-close kinetics exhibited double exponential behaviour, with a strong voltage dependence of the slow open time constant. Infrequently also a substate conductance level was identified. The voltage and calcium dependence suggest that the channel plays a role in adjusting K+ secretion to Na+ absorption in the fine regulation of cation excretion in renal collecting ducts.
    Type of Medium: Electronic Resource
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  • 4
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    Boulder : Periodicals Archive Online (PAO)
    East European quarterly. 22:1 (1988:Spring) 1 
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