Many members of the synaptotagmin (Syt) protein family bind Ca(2+) and trigger exocytosis, but some Syt proteins appear to have no Ca(2+)-dependent actions and their biological functions remain obscure. Syt IV is an activity-induced brain protein with no known Ca(2+)-dependent interactions and its subcellular localization and biological functions have sparked considerable controversy. We found Syt IV on both micro- and dense-core vesicles in posterior pituitary nerve terminals in mice. In terminals from Syt IV knockout mice compared with those from wild types, low Ca(2+) entry triggered more exocytosis, high Ca(2+) entry triggered less exocytosis and endocytosis was accelerated. In Syt IV knockouts, dense-core and microvesicle fusion was enhanced in cell-attached patches and dense-core vesicle fusion pores had conductances that were half as large as those in wild types. Given the neuroendocrine functions of the posterior pituitary, changes in Syt IV levels could be involved in endocrine transitions involving alterations in the release of the neuropeptides oxytocin and vasopressin.