1. Perezone (PZN) on the in vitro intestinal smooth muscle of the rat relaxes the basal tonus of the smooth muscle, interrupts spontaneous activity and also blocks the contractile response induced by ACh, K+, and Ba2+; these actions are dose dependent. 2. Although in presence of small doses of PZN, the isometric contractile response to ACh was increased. 3. In calcium free intestinal smooth muscle preparation, the addition of PZN in low dose before Ca2+ increased the contractile effect of added calcium to the bath, but in presence of high doses of PZN the response to calcium was depressed. 4. PZN in calcium free preparations antagonized the contraction caused by adding barium. 5. These findings suggested that with small doses of PZN more availability of intracellular calcium free exist and/or an increase in excitability and/or an inhibition of AChase could coexist. 6. The responses of the intestine to high doses of PZN were possibly in part by blocking calcium entry. 7. The smooth muscle responses to PZN suggest that it has a membranal effect and/or an action on the internal calcium stores possibly increasing the intracellular calcium concentration. It is likely to be the expression of an increase in the intracellular calcium concentration above the levels normally reached that would be responsible for uncoupling of the smooth muscle, which would occur if the [Ca2+]i rises excessively.