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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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