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Am J Physiol Renal Physiol 281: F687-F692, 2001;
0363-6127/01 $5.00
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Vol. 281, Issue 4, F687-F692, October 2001

Modulation of aldosterone-induced stimulation of ENaC synthesis by changing the rate of apical Na+ entry

Lisette Dijkink, Anita Hartog, Carel H. Van Os, and René J. M. Bindels

Department of Cell Physiology, University Medical Centre Nijmegen, 6500 HB Nijmegen, The Netherlands

Primary cultures of immunodissected rabbit connecting tubule and cortical collecting duct cells were used to investigate the effect of apical Na+ entry rate on aldosterone-induced transepithelial Na+ transport, which was measured as benzamil-sensitive short-circuit current (Isc). Stimulation of the apical Na+ entry, by long-term short-circuiting of the monolayers, suppressed the aldosterone-stimulated benzamil-sensitive Isc from 320 ± 49 to 117 ± 14%, whereas in the presence of benzamil this inhibitory effect was not observed (335 ± 74%). Immunoprecipitation of [35S]methionine-labeled beta -rabbit epithelial Na+ channel (rbENaC) revealed that the effects of modulation of apical Na+ entry on transepithelial Na+ transport are exactly mirrored by beta -rbENaC protein levels, because short-circuiting the monolayers decreased aldosterone-induced beta -rbENaC protein synthesis from 310 ± 51 to 56 ± 17%. Exposure to benzamil doubled the beta -rbENaC protein level to 281 ± 68% in control cells but had no significant effect on aldosterone-stimulated beta -rbENaC levels (282 ± 68%). In conclusion, stimulation of apical Na+ entry suppresses the aldosterone-induced increase in transepithelial Na+ transport. This negative-feedback inhibition is reflected in a decrease in beta -rbENaC synthesis or in an increase in beta -rbENaC degradation.

rabbit kidney; cortical collecting duct; connecting tubule; epithelial sodium channel; benzamil


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