Expression of amiloride-blockable sodium channels in Xenopus oocytes.
This report describes the expression of a sodium-selective, amiloride-blockable conductance in Xenopus oocytes that have been injected with RNA prepared from a distal nephron cell line (A6). After injecting the RNA into mature oocytes (stage V or VI) and incubating the oocytes for 2-4 days, the oocytes were examined for amiloride-blockable current. The RNA induced a substantial amiloride-blockable current. Uninjected or water-injected oocytes had no measurable amiloride-blockable current. RNA prepared from aldosterone-treated A6 cells was much more effective in inducing amiloride-blockable sodium current than RNA prepared from aldosterone-depleted A6 cells. Oocytes injected with RNA prepared from mineralocorticoid-depleted cells appeared very similar to water-injected oocytes. The amiloride-blockable current in oocytes has a reversal potential of approximately +50 - +60 mV, which varies 61 mV/decade change in external sodium concentration, suggesting that the current is highly selective for sodium over other ions. In addition, the concentration of amiloride that produces half block of the current is 48 +/- 8 nM. Thus the current expressed in oocytes appears very similar to sodium-selective currents observed from the apical membranes of various tight epithelial tissues.
Hinton
C F
CF
Department of Physiology, Emory University Medical School, Atlanta, Georgia 30322.
Eaton
D C
DC
D000584:Amiloride / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D005260:Female; D008564:Membrane Potentials / Q000187:drug effects; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics*; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D015222:Sodium Channels / Q000187:drug effects / Q000502:physiology*; D014982:Xenopus laevis
D008565:Membrane Proteins; D015222:Sodium Channels; D012313:RNA; D000584:Amiloride