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  • 1
    ISSN: 1432-1424
    Keywords: Active transport ; oxidative metabolism ; toad bladder
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The use of an Ussing chamber with well-defined mixing characteristics coupled to a mass spectrometer permits the concurrent evaluation of transepithelial current and oxidative metabolism with improved temporal resolution. The time-course of the amiloride-sensitive currentI a and the rate of suprabasal CO2 productionJ CO2 sb were observed in 10 toad urinary bladders at short-circuit and after clamping ΔΨ at 100 mV, serosa positive. Following perturbation of ΔΨ (0→100mV),I a declined sharply within 1/2 min, remained near constant ∼15 min, and then increased slightly.J CO2 sb declined more gradually, remained near constant at ∼4–7 min, and then declined further. Detailed analysis revealed an early quasi-steady state with near constancy ofJ CO2 sb starting at 2.9±1.1 (sd) min and lasting 4.7 ±1.8 (sd) min, followed by relaxation to a later steady state at about 15 min. During the early quasi-steady state,I a was also nearly constant. Considering that in steady statesI a/F≃J Na a , the rate of transepithelial active Na transport, during the early quasi-steady state mean values ±se ofJ Na a ,J CO2 sb and (J Na a /J CO2 sb ) were, respectively, 29.9±1.7%, 59.4 ±3.2%, and 56.4±5.7% of values at short-circuit. Corresponding values during the late steady state were 41.4±6.0%, 38.2±6.1%, and 111.3±8.6%. Thus the flow ratioJ Na a /J CO2 sb was depressed significantly during the early quasi-steady state, but returned later to the original value. The results of measurements ofI a andJ CO2 sb in three hemibladders were qualitatively similar. In terms of a phenomenological “black-box” treatment the findings are consistent with earlier studies indicating incomplete coupling between transport and metabolism. Further studies will be required to clarify the molecular basis for these observations.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 40 (1978), S. 15-27 
    ISSN: 1432-1424
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Active sodium transport is classically analyzed in terms of an equivalent circuit, comprising an active conductancek a and an electromotive force of sodium transportE Na. AlthoughE Na is commonly considered the driving force of transport, model experiments have suggested thatE Na is a composite parameter, incorporating both kinetic and energetic factors. An alternative approach considers both transport and the associated oxidative metabolism in terms of a nonequilibrium thermodynamic (NET) formulation, involving phenomenological coefficients and the affinityA, presumed to represent kinetic and energetic factors, respectively. Model experiments testing the NET formulation suggest that the affinity is indeed an energetic parameter. Calculated values ofA in untreated frog skins and toad bladders range from about 20 to 80 kcal per mole of O2 consumption. Assuming a P/O ratio of 3, this range corresponds to about 3–13 kcal per mole of ATP utilization, values compatible with reported direct measurements. Although brief perturbations of transepithelial electrical potential ΔΨ resulted in linear current-voltage relationships, indicating constancy ofE Na andk a , 15-min perturbations of ΔΨ resulted in nonlinearity, indicating changes inE Na andk a : perturbations of ΔΨ enhancing active transport were associated with decrease ofE Na and increase ofk a ; slowing of active transport produced the converse effects.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-1424
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 69 (1982), S. 125-136 
    ISSN: 1432-1424
    Keywords: membrane potentials ; membrane conductances ; frog skin ; constant field equation ; sodium conductances
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Previous studies in anuran epithelia have shown that, after clamping the transepithelial voltage in symmetrical sequences for 4–6 min there is near-constancy of the rate of active Na transport and the associated oxidative metabolism, with a near-linear potential dependence of both. Here we have investigated in frog skin the cellular electrophysiological events associated with voltage clamping (V t =inside-outside potential). Increase and decrease ofV t produced converse effects, related directly to the magnitude ofV t . Hyperpolarization resulted in prompt decrease in inward transepithelial currentI t and increase in fractional outer membrane resistancefR 0 (as evaluated from small transient voltage perturbations) and in outer membrane potentialV 0. Overshoot ofV 0 was followed by relaxation to a quasi-steady state in minutes. Changes infR 0 were progressive, with half times of some 1–5 sec. Changes in transepithelial slope conductanceg t were more variable, usually preventing precise evaluation of the outer and inner cell membrane conductancesg 0 andg i . Nevertheless, it was shown thatg 0 is related inversely toV t andV 0. Presuming insensitivity ofV i toV t , the dependence ofg 0 onV 0 in the steady state much exceeds that predicted by the constant field equation. Apparent inconsistencies with earlier results of others may be attributable to differences in protocol and the complex dependence ofg 0 onV 0 and/or cellular current. In contrast to previous findings in tight epithelia at open circuit, differences inV t were associated with substantial differences infR 0 and inner membrane potentialV i . Hyperpolarization ofV t over ranges commonly employed in studies of active transport and metabolism appears to increase significantly the electrochemical work per Na ion transported.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-1424
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 53 (1980), S. 235-236 
    ISSN: 1432-1424
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 55 (1980), S. 53-68 
    ISSN: 1432-1424
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Determinations of current-voltage relationships are widely employed in the characterization of epithelial sodium transport. In order to determine the protocol dependence of transport parameters in the toad urinary bladder, studies were carried out in the presence and absence of amiloride, an inhibitor of active sodium transport. With symmetric positive and negative perturbations of the transepithelial electrical potential difference Δψ (0→±100 mV) for 30 sec, the amiloride-sensitive current-voltage (i a -Δψ) relationship was near linear over the range −75→+100 mV, indicating constancy of the conductance κa and the apparent electromotive force “E Na”, lumped parameters of the standard electrical equivalent circuit model of the active transport system. With a reverse protocol (±100→0 mV) or 15 min perturbations thei a -Δψ relationships were highly nonlinear. Nonlinearity reflected voltage dependence of parameters: perturbations that increased active transport decreased “E Na” and increased κa, as evaluated from 10 sec perturbations of Δψ; slowing of active transport produced the converse changes. These effects are usefully analyzed in both quasi-steady states and true steady states by means of a detailed equivalent circuit incorporating the significant ionic currents across each plasma membrane. Precise understanding of the significance of κa and “E Na” will require characterization of the partial ionic conductances on perturbation of Δψ.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 50 (1979), S. 205-224 
    ISSN: 1432-1424
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The flows of solute molecules in a membrane under the influence of concentration gradients are considered within the framework of classical physical theories. A lattice model is constructed in which the binding sites represent potential minima and the flows are regarded as a result of molecules' making discrete transitions between the binding sites. Expressions for two-component currents are derived from certain descriptions for the transition mechanism. Where the molecular movement is given the crudest description, permeability coefficients are identical for both components and there is no current coupling. Where the molecular movement is given some finer detail, the permeability coefficients differ and positive coupling of flows appears. Our result applies to a combination of flows of tracer and abundant species as well as, more generally, to any combination of flows of two components which are distinguishable yet kinetically similar. Also considered are binary currents whose transport mechanism is further controlled by allosteric cooperativity. Whether the cooperative control is short or long ranged, permeability coefficients and fluxes differ appreciably from those without cooperative control. Thus, unlike in the case of channel flow, current coupling here may be either positive or negative, depending on the strength and nature of cooperative coupling. Numerical evidence suggests that the permeability and coupling may have discontinuous behavior, possibly indicating the existence of phase transitions. Our lattice model, from which the formulations for the flows are obtained, is compatible with current concepts of membrane structure.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 197 (1963), S. 701-701 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] To explain these findings in frog skin, Koefoed-Johnsen and Ussing have proposed a model which, as applied to the toad bladder, presumes passive entry of sodium through a selective permeability barrier at the mucosal surface, followed by active transport by a 'sodium pump' at the serosal surface6. ...
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Pflügers Archiv 399 (1983), S. 336-341 
    ISSN: 1432-2013
    Keywords: Frog skin ; Junctional conductance ; Voltagedivider ratio ; Paracellular conductance ; Membrane conductances
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract It has been suggested that distribution of lateral interspace resistance in association with a highly conductive junction can significantly affect the measurement of outer membrane(o)/epithelial(t) voltage divider ratios (F o=δV o/δV t), thereby leading to erroneous inferences regarding the outer membrane fractional resistance [fR o=R o/R c=R o/(R o+R i)], whereR o andR i are the outer and inner cell membrane resistance respectively andR c is the total cell membrane resistance. We present here experimental evidence for this point of view. During seasons when frog skins were highly permeable to Cl, transepithelial conductanceg t often exceeded 2 mS/cm2. High concentrations of external amiloride rapidly blocked cellular transport, butg t initially remained high andF o remained appreciably less than 1.0. These values ofF o were found here to result from low junctional resistanceR j: increase ofR j, either gradually following the administration of amiloride, or abruptly with external replacement of Cl by other anions, was associated with increase ofF o to near unity, without effect on the membrane potential or significant change in the short-circuit current. Experimental results following amiloride validated a simple equivalent circuit model predicting near-linear increase inF o with progressive decrease ing t and led to plausible values ofR j and lateral space resistanceR l. The possible influence of the paracellular resistance pattern on the evaluation of cell membrane resistances from voltage divider ratios is discussed.
    Type of Medium: Electronic Resource
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