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Am J Physiol Cell Physiol 266: C1513-C1522, 1994;
0363-6143/94 $5.00
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AJP - Cell Physiology, Vol 266, Issue 6 C1513-C1522, Copyright © 1994 by American Physiological Society

ARTICLES

Antidiuretic hormone-responding nonselective cation channel in distal nephron epithelium (A6)

Y. Marunaka, H. Tohda, N. Hagiwara and T. Nakahari
Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.

Arginine vasotocin (AVT, 70 mU/ml) added from the basolateral side transiently activated a nonselective cation (NSC) channel with a single-channel conductance of 28.5 pS and almost identical selectivity for Na+ and K+ in the apical membrane of distal nephron cells (A6) cultured on permeable supports for 10-12 days in media containing 10% fetal bovine serum without supplemental aldosterone. The open probability (Po) of the NSC channel at the apical resting membrane potential in cell-attached patches was approximately 0.09 and increased when the apical membrane depolarized. The Po of the NSC channel was decreased by a rise in cytosolic Ca2+ concentration within a range of 30 nM-1 microM but not affected by cytosolic pH within a range of 6-8. The channel was activated by the application of negative pressure (10-60 cmH2O) into the patch pipette. Gadolinium (2 microM), an inhibitor of stretch-activated channels, decreased the Po by 40%. This blocking action of gadolinium was more effective after the channel was activated by stretch, i.e., 2 microM gadolinium decreased the Po by 70% when a negative pressure (60 cmH2O) was applied into the patch pipette. Amiloride (10 and 100 microM) showed a blocking action on the channel only when the NSC channel was activated by stretch.


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