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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 106 (1988), S. 13-28 
    ISSN: 1432-1424
    Keywords: cell potential ; amiloride ; sodium transport ; reversal potential
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Knowledge of the voltage dependencies of apical and basolateral conductances is important in determining the factors that regulate transcellular transport. To gain this knowledge it is necessary to distinguish between cellular and paracellular currents and conductances. This is generally done by sequentially measuring transepithelial current/voltage (I t /V t ) and conductance/voltage (g t /V t ) relationships before and after the abolition of cellular sodium transport with amiloride. Often, however, there are variable time-dependent and voltage-dependent responses to voltage perturbation both in the absence and presence of amiloride, pointing to effects on the paracellular pathway. We have here investigated these phenomena systematically and found that the difficulties were significantly lessened by the use of an intermittent technique, measuringI t andg t before and after brief (〈10 sec) exposure to amiloride at each setting ofV t .I/V relationships were characterized by these means in frog skins (Rana pipiens, Northern variety, andRana temporaria). Cellular current,I c , decreased with hyperpolarization (larger serosa positive clamps) ofV t . DerivedI c /V t relationships betweenV t =0 and 175 mV (serosa positive) were slightly concave upwards. Because values of cell conductance,g c , remained finite, it was possible to demonstrate reversal ofI c . Values of the reversal potentialV' averaged 156±14 (sd,n=18) mV. Simultaneous microelectrode measurements permitted also the calculation of apical and basolateral conductances,g a andg b . The apical conductance decreased monotonically with increasing positivity ofV t (andV a ). In contrast, in the range in which the basolateral conductance could be evaluated adequately (V t 〈125 mV),g b increased with more positive values ofV t (andV b ). That is, there was an inverse relation betweeng b and cellular current at the quasi-steady state, 10–30 sec after the transepithelial voltage step.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 61 (1981), S. 127-134 
    ISSN: 1432-1424
    Keywords: Frog skin ; microelectrodes ; membrane potentials ; intracellular activities ; amiloride
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Intracellular Na+, K+, and Cl− activities (a Na i ,a K i ,a Cl i ) and transapical membrane potentials (V o) were measured with liquid ion-exchanger and open-tip microelectrodes in isolated short-circuited frog skins (R. pipiens) incubated at 23°C in normal amphibian Ringer's solution. Under control conditionsa Na i =14±3mm,a K i =132±10mm anda Cl i =18±3mm (sd). The value ofa Cl i is 4.4 times the value corresponding to electrochemical equilibrium for this ion. Thus, Cl− is actively accumulated by epithelial cells of the frog skin. Shortly after addition of amiloride (2–5 μm) to the apical bathing medium,a K i ,a Na i , anda Cl i were essentially unchanged althoughV o had hyperpolarized by about 30–40 mV. During long-term exposure to amiloridea K i anda Cl i did not change significantly,V o depolarized by about 16 mV from the maximal value anda Na i decreased to 8±3mm. Immediately after exposure to amiloride the transmembrane driving force for Na+ increased from 124 to 154 mV. During further exposure to amiloride, despite changes in bothV o anda Na i , this driving force remained virtually constant. SinceI sc during this period was close to zero, it is suggested that the observed driving force for Na+ under these conditions approximates the maximal driving force generated by the Na+−K+ ATP-ase pump in the basolateral cell membrane.
    Type of Medium: Electronic Resource
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  • 3
    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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