Calcium flux across organelle and plasma membranes is an important event in neutrophil activation. We measured calcium discharge into the media from neutrophils stimulated with formyl-methionyl-leucyl-phenylalanine after treatment with cytochalasin b. Cytochalasin markedly potentiated calcium efflux from stimulated neutrophils, and similarly promoted release of lysosomal enzymes into the media. Colchicine neither reproduced nor modified the cytochalasin effect. Neutrophil cytoplasts discharged very little calcium in response to stimulation, and discharge was not significantly altered by cytochalasin b. These findings indicate that neutrophil degranulation is accompanied by efflux of calcium into the media, and suggest that the neutrophil granules constitute a source of mobilizable calcium which could be used to modify the extracellular microenvironment.