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Significance Neuromodulators are released by most neurons and control many brain functions, but their release mechanism remains largely elusive. Calcium sensors for other secretory pathways have been characterized but not for neuromodulator release in the brain. Using release assays with single-vesicle resolution in living neurons, we identify the sensors that drive neuromodulator release from dense core vesicles. Neuromodulator release depends equally on two sensors, Syt1 and Syt7, that are both rate limiting for release and compensate for each other upon overexpression (cross-rescue), despite being in different locations in the cell. These data show that the mechanism of neuromodulator secretion is different from previously studied systems.

Neuromodulator release in neurons requires two functionally redundant calcium sensors