Current-noise analysis of Na absorption in the embryonic coprodeum: stimulation by aldosterone and thyroxine.
The mechanism and regulation of sodium transport in the embryonic coprodeum of chicken were investigated with isolated epithelia in vitro by electrophysiological techniques. Electrogenic sodium transport (INa) was measured in Ussing chambers by the short-circuit current (Isc) technique and identified by the diuretic amiloride or by removal of sodium from the apical medium. Apical sodium channels and the kinetics of amiloride binding were investigated by current-noise analysis. Isc and INa were measured under control conditions and under the influence of in vitro incubation with aldosterone and thyroxine. At 20 days the embryonic coprodeum has an Isc of 12.6 +/- 1.4 microA/cm2 and a transepithelial resistance of 519 +/- 40 omega.cm2. Amiloride blocks 9.0 +/- 1.3 microA/cm2 of the Isc, which represents electrogenic Na+ absorption and can be inhibited by serosal ouabain. Aldosterone does not stimulate Isc or INa, whereas thyroxine increases Isc and INa about threefold. Aldosterone in combination with thyroxine increases Isc and INa further to about five- to sixfold. In both cases the hormonal stimulation can be totally blocked by spironolactone. Current-noise analysis of the apical Na+ entry step reveals amiloride-sensitive Na+ channels with a single-channel current of approximately 2.3 pA and a channel density of 9-16 million/cm2 under stimulated conditions. Half-maximal amiloride block occurs at 0.8-1 microM. The hormones stimulate Na+ absorption by increasing the Na+ channel density and not the single-channel current.(ABSTRACT TRUNCATED AT 250 WORDS)
Clauss
W
W
Institut für Tierphysiologie, Justus-Liebig-Universität Giessen, Germany.
Hoffmann
B
B
Krattenmacher
R
R
Van Driessche
W
W
D000042:Absorption; D000450:Aldosterone / Q000494:pharmacology*; D000584:Amiloride / Q000494:pharmacology; D000818:Animals; D002642:Chick Embryo; D002988:Cloaca / Q000187:drug effects / Q000196:embryology / Q000502:physiology*; D004553:Electric Conductivity / Q000187:drug effects; D004594:Electrophysiology / Q000379:methods; D004848:Epithelium / Q000502:physiology; D007408:Intestinal Absorption / Q000187:drug effects*; D015640:Ion Channel Gating / Q000187:drug effects; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology; D008433:Mathematics; D008954:Models, Biological; D010042:Ouabain / Q000494:pharmacology; D012964:Sodium / Q000378:metabolism*; D013148:Spironolactone / Q000494:pharmacology; D013974:Thyroxine / Q000494:pharmacology*
D007473:Ion Channels; D013148:Spironolactone; D000450:Aldosterone; D010042:Ouabain; D000584:Amiloride; D012964:Sodium; D013974:Thyroxine