AJP - Cell Physiology, Vol 270, Issue 1 C224-C234, Copyright © 1996 by American Physiological Society

ARTICLES

Stretch modulates amiloride sensitivity and cation selectivity of sodium channels in human B lymphocytes

J. M. Achard, J. K. Bubien, D. J. Benos and D. G. Warnock
Department of Medicine, University of Alabama at Birmingham 35294-0007, USA.

Stretch-mediated regulation of amiloride-sensitive Na+ channels was examined in Epstein-Barr virus-transformed human B lymphocytes. Cation conductances were measured using whole cell patch-clamp techniques. Stretch activation, induced by increasing the hydrostatic pressure of the bath solution, immediately and reversibly increased both inward and outward ionic conductances once a threshold of 2-5 mmH2O was reached. Ionic substitutions confirmed that stretch enhanced membrane conductivity for both Na+ and K+. Amiloride (2 microM) completely prevented the response to elevated hydrostatic pressure; however, when amiloride was applied after stretch-induced activation, the sensitivity to amiloride was dramatically decreased (inhibitor concentration that reduces whole cell current by 50% of approximately 20 microM). Evidence that the currents induced by stretch were mediated by Na+ channels was provided by the lack of response to stretch in lymphocytes from patients with Liddle's syndrome, which is caused by expression of a truncated mutant of the beta-subunit of the amiloride-sensitive Na+ channel. Pretreatment with colchicine (0.5 mM, 30 min) prevented stretch-induced activation, which shows evidence of the involvement of the cytoskeleton. These data indicate that stretch regulates the conductance of amiloride-sensitive Na+ channels in immortalized human B lymphocytes and also alters its cationic selectivity and its sensitivity to amiloride.

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