Selective expression of an amiloride-inhibitable Na+ conductance from mRNA of respiratory epithelium in Xenopus laevis oocytes.
Poly(A)+RNA was prepared from primary cultures of human nasal polyp epithelia and from native bovine tracheal epithelia. Six to fifty nanograms mRNA were injected into prophase-arrested immature Xenopus laevis oocytes. One to four days later the oocytes were probed with electrophysiological techniques for induction of novel ion conductances. Oocytes injected with mRNA had lower membrane potentials (Vm) and resistances (Rm) than controls. By use of step changes in extracellular Na+ concentration and applying amiloride, a Na+ conductance could be identified in mRNA-injected oocytes, which was inhibited by submicromolar concentrations of amiloride with the same kinetics (Ki = 1.3 X 10(-7) mol/l) as in the original tissue. After complete inhibition of this conductance by 10(-5) mol/l amiloride, Vm and Rm approached the respective values of controls. The data indicate that oocytes express functional epithelial Na+ channels but apparently no other epithelial ion conductances from injected mRNA of respiratory epithelium.
Kroll
B
B
Zentrum der Physiologie, Klinikum der J.W.-Goethe-Universität, Frankfurt, Federal Republic of Germany.
Bautsch
W
W
Bremer
S
S
Wilke
M
M
Tümmler
B
B
Frömter
E
E
D000584:Amiloride / Q000494:pharmacology*; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004594:Electrophysiology / Q000379:methods; D004848:Epithelium / Q000378:metabolism; D005260:Female; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D008565:Membrane Proteins / Q000235:genetics*; D009298:Nasal Polyps / Q000378:metabolism*; D009865:Oocytes / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics* / Q000302:isolation & purification; D015222:Sodium Channels / Q000187:drug effects / Q000502:physiology*; D014982:Xenopus laevis
D008565:Membrane Proteins; D012333:RNA, Messenger; D015222:Sodium Channels; D000584:Amiloride