Synaptotagmin I (Syt I), a possible Ca 2+ sensor for neurotransmitter release, was suggested to be involved in neurite outgrowth of chick dorsal root ganglion (DRG) neurons, based on introduction of the antibody against the C2A domain into cells via mechanical lesions. Recently, however, the functional block antibody against the C2A domain was shown to block axonal repair processes, which raised a question as to whether Syt I is indeed involved in neurite outgrowth. In this study, we expressed Syt I or II in PC12 cells and found that these expression did indeed promote neurite outgrowth, as compared to control cells. We further showed that expression of the phospholipid binding activity-deficient mutant of Syt II (Δ180–183) had little effect on the neurite outgrowth of PC12 cells. These results indicate the Ca 2+/phospholipid binding site of Syt I or II to be essential for neurite outgrowth.