Steroid hormone-dependent expression of blocker-sensitive ENaCs in apical membranes of A6 epithelia.
Weak channel blocker-induced noise analysis was used to determine the way in which the steroids aldosterone and corticosterone stimulated apical membrane Na+ entry into the cells of tissue-cultured A6 epithelia. Among groups of tissues grown on a variety of substrates, in a variety of growth media, and with cells at passages 73-112, the steroids stimulated both amiloride-sensitive and amiloride-insensitive Na+ transport as measured by short-circuit currents in chambers perfused with either growth medium or a Ringer solution. From baseline rates of blocker-sensitive short-circuit current between 2 and 7 microA/cm2, transport was stimulated about threefold in all groups of experiments. Single channel currents averaged near 0.3 pA (growth medium) and 0.5 pA (Ringer) and were decreased 6-20% from controls by steroid due to the expected decreases of fractional transcellular resistance. Irrespective of baseline transport rates, the steroids in all groups of tissues stimulated transport by increase of the density of blocker-sensitive epithelial Na+ channels (ENaCs). Channel open probability was the same in control and stimulated tissues, averaging approximately 0.3 in all groups of tissues. Accordingly, steroid-mediated increases of open channel density responsible for stimulation of Na+ transport are due to increases of the apical membrane pool of functional channels and not their open probability.
Baxendale-Cox
L M
LM
Department of Molecular and Integrative Physiology, University of Illinois at Urbana-Champaign 61801, USA.
Duncan
R L
RL
Liu
X
X
Baldwin
K
K
Els
W J
WJ
Helman
S I
SI
D000450:Aldosterone / Q000494:pharmacology*; D000584:Amiloride / Q000031:analogs & derivatives / Q000494:pharmacology*; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division; D002460:Cell Line; D003345:Corticosterone / Q000494:pharmacology*; D003470:Culture Media; D004553:Electric Conductivity; D004847:Epithelial Cells / Q000378:metabolism; D053503:Epithelial Sodium Channels; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects; D011336:Probability; D026941:Sodium Channel Blockers; D015222:Sodium Channels / Q000096:biosynthesis / Q000502:physiology*
D003470:Culture Media; D053503:Epithelial Sodium Channels; D026941:Sodium Channel Blockers; D015222:Sodium Channels; C058206:6-chloro-3,5-diaminopyrazine-3-carboxamide; D000450:Aldosterone; D000584:Amiloride; D003345:Corticosterone