Acetylcholine and caffeine activate cl and suppress k+ conductances in human bronchial smooth muscle. Janssen, Luke J. Department of Medicine, McMaster University, Hamilton, Ontario, Canada L8N 3Z5
APStracts 2:0224L, 1995.
The conductance changes underlying agonist-evoked depolarization in human airway smooth muscle (ASM) were examined using single ASM cells liberated enzymatically from non-carcinomatous bronchi and studied using patch-clamp techniques. Step commands to potentials at or more positive than the resting membrane potential evoked outward current, which was predominantly delayed rectifier K+ current with some Ca2+ -dependent K+ current. Caffeine (5 mM) evoked depolarization and contraction lasting several minutes. During voltage clamp at -60 mV, caffeine evoked inward current with a latency of nearly equal to 1 s, mean amplitude of 320+/-65 pA, and a duration of nearly equal to 5 s (even though agonist application exceeded this duration). Using the perforated-patch configuration, these responses could be evoked repeatedly at 4 minute intervals for up to 30 minutes; rupture of the membrane and dialysis of the cytosol, however, abrogated the responses to caffeine. The current was outwardly rectifying with mean reversal potential (Vrev) of -31+/-4 mV. When K+ conductances were blocked by Cs+, the current-voltage relationship was linear (i.e. an outwardly-rectifying component was eliminated) and Vrev was displaced in the positive direction to +2+/-1 mV. Changes in the Cl_ equilibrium potential were accompanied by a displacement of Vrev in a manner predicted by the Nernst equation for a Cl_ current. The effects of caffeine were mimicked by acetylcholine; in addition, acetylcholine and caffeine each occluded the response to the other agonist. Spasmogens also caused a prolonged suppression of K+ currents (both Ca2+-dependent and delayed rectifier). We conclude that, in human ASM, acetylcholine and caffeine cause a transient activation of Ca2+-dependent Cl_ current (due to release of internal Ca2+) and prolonged suppression of K+ currents, leading to depolarization and contraction.

Received 6 April 1995; accepted in final form 4 December 1995.
APS Manuscript Number L108-5.
Article publication pending Am. J. Physiol. (Lung Cell. Mol.
Physiology).
ISSN 1080-4757 Copyright 1995 The American Physiological Society.
Published in APStracts on 12 December 95