The relationship of CaZ+ and plasmodesmatal closure was examined in staminal hairs of Setcreasea purpurea by microinjecting cells with active mastoparan (Mas-7), inactive mastoparan (Mas-1 7), active inositol-1,4,5-trisphosphate (IP,), or inactive I P, . Calcium green dextran 10,000 was used to study cellular free Caz+, and carboxyfluorescein was used to monitor plasmodesmatal closure.When Mas-7 was microinjected into the cytoplasm of cell 1 (the tip cell of a chain of cells), a rapid increase i n calcium green dextran-10,000 fluorescence was observed in the cytoplasmic areas on both sides of the plasmodesmata connecting cells 1 and 2 during the same time that the diffusion of carboxyfluorescein through them was blocked. The inhibition of cell-to-cell diffusion was transient, and the closed plasmodesmata reopened within 30 s. The elevated Ca2+ level near plasmodesmata was also transient and returned t o base level in about 1.5 min. The transient increase in CaZ+, once initiated in cell 1, repeated with an oscillatory period of 3 min. Elevated Caz+ and oscillations of Caz+ were also observed near interconnecting cell walls throughout the chain of cells, indicating that the signal had been transmitted. Previously, we reported that IP, closed plasmodesmata; now we report that it stimulated Ca2+ and oscillations similar to Mas-7. The effect was specific for similar concentrations of Mas-7 over Mas-1 7 and active IP, over inactive IP,. It is important that the Ca2+ channel blocker La3+ eliminated the responses from Mas-7 and I P, , indicating that an influx of Ca2+ was required. These results support the contention that plasmodesmata functioning is regulated via Ca2+ and that IP, may be an intermediary between the stimulus and CaZ+ elevations.