1. Acetylcholine (0.1 mmol/l) and KCl (80 mmol/l) induce contractile responses in guinea-pig trachea. 2. In the absence of extracellular Ca, contractile responses to ACh or KCl decrease as the preincubation time or the EDTA concentration increases. 3. Exposure of the tissue to Ca-free medium decreases the basal tone. 4. Prolonged incubation in the presence of EDTA (1 mmol/l) promotes a delay in the recovery of the tonic component of the contraction elicited subsequently by acetylcholine and KCl in physiological salt solution, and abolishes the phasic one.