Differential Co-release of Two Neurotransmitters from a Vesicle Fusion Pore in Mammalian Adrenal Chromaffin Cells

Co-release of multiple neurotransmitters from secretory vesicles is common in neurons and neuroendocrine cells. However, whether and how the transmitters co-released from a single vesicle are differentially regulated remains unknown. In matrix-containing dense-core vesicles (DCVs) in chromaffin cells, there are two modes of catecholamine (CA) release from a single DCV: quantal and sub-quantal. By combining two microelectrodes to simultaneously record co-release of the native CA and ATP from a DCV, we report that (1) CA and ATP were co-released during a DCV fusion; (2) during kiss-and-run (KAR) fusion, the co-released CA was sub-quantal, whereas the co-released ATP was quantal; and (3) knockdown and knockout of the DCV matrix led to quantal co-release of both CA and ATP even in KAR mode. These findings strongly imply that, in contrast to sub-quantal CA release in chromaffin cells, fast synaptic transmission without transmitter-matrix binding is mediated exclusively via quantal release in neurons. •One vesicle releases two native transmitters via two modes (quantal versus sub-quantal)•Sub-quantal is produced jointly by fusion pore and intravesicular matrix•Co-release of catecholamine and ATP from a 100-nm vesicle recorded by two electrodes Zhang et al. uncover the mystery of sub-quanta and quanta during vesicular release of multi-neurotransmitters. Catecholamine sub-quanta are jointly produced by fusion pores and the vesicular matrix, whereas the co-released ATP quanta are fully released during transient fusion events.