Amiloride-sensitive Na+ transport across cultured renal (A6) epithelium: evidence for large currents and high Na:K selectivity.
Electrical techniques were used to determine the Na:K selectivity of the amiloride-sensitive pathway and to characterize cellular and paracellular properties of A6 epithelium. Under control conditions, the mean transepithelial voltage (VT) was -57 +/- 5 mV, the short-circuit current (Isc) averaged 23 +/- 2 microA/cm2 and the transepithelial resistance (RT) was 2.8 +/- 0.3 k omega cm2 (n = 13). VT and Isc were larger than reported in previous studies and were increased by aldosterone. The conductance of the amiloride-sensitive pathway (Gamil) was assessed before and after replacement of Na+ in the mucosal bath by K+, using two independent measurements: (1) the slope conductance (GT), determined from current-voltage (I-V) relationships for control and amiloride-treated tissues and (2) the maximum amiloride-sensitive conductance (Gmax) calculated from the amiloride dose-response relationship. The ratio of Gamil in mucosal Na+ solutions to Gamil for mucosal K+ solutions was 22 +/- 6 for GT measurements and 15 +/- 2 for Gmax data. Serosal ion replacements in tissues treated with mucosal nystatin indicated a potassium conductance in the basolateral membrane. Equivalent circuit analyses of nystatin and amiloride data were used to resolve the cellular (Ec) and paracellular (Rj) resistances (approximately 5 k omega cm2 and 8-9 k omega cm2, respectively). Analysis of I-V relationships for tissues depolarized with serosal K+ solutions revealed that the amiloride-sensitive pathway could be described as a Na+ conductance with a permeability coefficient (PNa) = 1.5 +/- 0.2 x 10(-6) cm/s and the intracellular Na+ concentration (Nai) = 5 +/- 1 mM (n = 5), similar to values from other tight epithelia. We conclude that A6 epithelia are capable of expressing large amiloride-sensitive currents which are highly Na+ selective.
Wills
N K
NK
Department of Physiology and Biophysics, University of Texas Medical Br., Galveston 77550-2781.
Millinoff
L P
LP
D000450:Aldosterone / Q000494:pharmacology; D000584:Amiloride / Q000494:pharmacology*; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002460:Cell Line; D004553:Electric Conductivity; D004848:Epithelium / Q000502:physiology; D007668:Kidney / Q000187:drug effects / Q000502:physiology*; D008564:Membrane Potentials / Q000187:drug effects; D009092:Mucous Membrane / Q000502:physiology; D009761:Nystatin / Q000494:pharmacology; D011188:Potassium / Q000378:metabolism; D015221:Potassium Channels / Q000187:drug effects / Q000502:physiology*; D012964:Sodium / Q000378:metabolism*; D015222:Sodium Channels / Q000187:drug effects / Q000502:physiology*; D014982:Xenopus laevis
D015221:Potassium Channels; D015222:Sodium Channels; D009761:Nystatin; D000450:Aldosterone; D000584:Amiloride; D012964:Sodium; D011188:Potassium