Intracellular ionic activities in frog skin

Abstract

Intracellular Na+, K+, and Cl- activities (aiNa, aiK, aiCl) and transapical membrane potentials (V0) were measured with liquid ion-exchanger and open-tip microelectrodes in isolated short-circuited frog skins (R. pipiens) incubated at 23 degrees C in normal amphibian Ringer's solution. Under control conditions aiNa = 14 +/- 3 mM, aiK = 132 +/- 10 mM and aiCl = 18 +/- 3 mM (SD). The value of aiCl 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 microM) to the apical bathing medium, aiK, aiNa, and aiCl were essentially unchanged although V0 had hyperpolarized by about 30--40 mV. During long-term exposure to amiloride aiK and aiCl did not change significantly, V0 depolarized by about 16 mV from the maximal value and aiNa decreased to 8 +/- 3 mM. 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 both V0 and aiNa, this driving force remained virtually constant. Since Isc during this period was close to zero, it is suggested that the observed driving force for Na+ under these condition approximates the maximal driving force generated by the Na+--K+ ATP-ase pump in the basolateral cell membrane.