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Asthma Research Group, Firestone Institute for Respiratory Health, St. Joseph's Hospital, Hamilton L8N 4A6; and the Department of Medicine, McMaster University, Hamilton, Ontario, Canada L8N 3Z5
We investigated the mechanisms
underlying muscarinic excitation-contraction coupling in canine airway
smooth muscle using organ bath, fura 2 fluorimetric, and patch-clamp
techniques. Cyclopiazonic acid (CPA) augmented the responses to
submaximal muscarinic stimulation in both tracheal (TSM) and bronchial
smooth muscles (BSM), consistent with disruption of the barrier
function of the sarcoplasmic reticulum. During maximal stimulation,
however, CPA evoked substantial relaxation in TSM but not BSM. CPA
reversal of carbachol tone persisted in the presence of
tetraethylammoium or high KCl, suggesting that hyperpolarization is not
involved; CPA relaxations were absent in tissues preconstricted with
KCl alone or by permeabilization with 
-escin, ruling out a
nonspecific effect on the contractile apparatus. Peak contractions were
sensitive to inhibitors of tyrosine kinase (genistein) or Rho kinase
(Y-27632). Sustained responses were dependent on Ca2+
influx in TSM but not BSM; this influx was sensitive to
Ni2+ but not La3+. In conclusion, there are
several mechanisms underlying excitation-contraction coupling in airway
smooth muscle, the relative importance of which varies depending on
tissue and degree of stimulation.
airway smooth muscle contraction; airway hyperresponsiveness; Rho kinase; tyrosine kinase; phosphatidylinositol 3-kinase; calcium ion-adenosinetriphosphatase; genistein; calcium ion-dependent chlorine ion current
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