ATP-receptors (P2X2, P2X3, P2X4 & P2X7) are found in neurohypophysial terminals (NHT). These purinergic receptor subtypes are known to be cation selective. Here we confirm that both sodium (Na+ ) and calcium (Ca2+ ) are permeable through these NHT purinergic receptors, but to varying degrees (91% vs. 9%, respectively). Furthermore, extracellular calcium inhibits the ATP-current magnitude. Thus, the objective of this study was to determine the effects of extracellular Na+ vs. Ca2+ on ATP-induced vasopressin (AVP) release from populations of rat isolated NHT. ATP (200 µM) perfused exogenously for 2 minutes in Normal Locke's buffer caused an initial transient increase in AVP release followed by a sustained increase in AVP release which lasted for the duration of the ATP exposure. Replacing extracellular NaCl with NMDG-Cl had no apparent effect on the ATP-induced transient increase in AVP release but abolished the sustained AVP release induced by ATP. Furthermore, removal of extracellular calcium resulted in no ATP-induced transient increase in AVP release, but had no effect on the delayed, sustained increase in AVP release. The ATP-induced calcium-dependent transient increase in AVP release was >95% inhibited by 10 µM of the P2X purinergic receptor antagonist PPADS, a dose sufficient to block P2X2 and P2X3 receptors but not P2X4 or P2X7 receptors. Interestingly, the ATP-induced calcium-independent, sodium-dependent sustained increase in AVP release was not affected by 10 µM PPADS. The ATP-induced calcium-dependent transient increase in AVP release was not affected by the P2X7 receptor antagonist BBG (100 nM). However, the ATP-induced sodium-dependent sustained AVP release was inhibited by 50%. Therefore, these results show that rat isolated NHT exhibit a biphasic response to exogenous ATP that is differentially dependent on extracellular calcium and sodium. Furthermore, the initial transient release appears to be through P2X2 and/or P2X3 receptors and the sustained release is through a P2X7 receptor. This article is protected by copyright. All rights reserved.