Autonomic neural activation of intracellular Ca²⁺ release in parotid acinar cells induces the secretion of the fluid and protein components of primary saliva critical for maintaining overall oral homeostasis. In the current study, we profiled the role of acidic organelles in shaping the Ca²⁺ signals of parotid acini using a variety of imaging and pharmacological approaches. Results demonstrate that zymogen granules predominate as an apically polarized population of acidic organelles that contributes to the initial Ca²⁺ release. Moreover, we provide evidence that indicates a role for the intracellular messenger NAADP in the release of Ca²⁺ from acidic organelles following elevation of cAMP. Our data are consistent with the "trigger" hypothesis where localized release of Ca²⁺ sensitizes canonical intracellular Ca²⁺ channels to enhance signals from the ER. Release from acidic stores may be important for initiating saliva secretion at low levels of stimulation and a potential therapeutic target to augment secretory activity in hypofunctioning salivary glands.