Neurexin mediates neuropeptide release from cholinergic motor neurons through dense-core vesicle localization

Neurexins are critical synaptic cell adhesion molecules that play many roles in modulating neurotransmitter release and synaptic function, and are high-confidence risk genes for neurodevelopmental conditions such as autism. Understanding the function of the neurexin superfamily has been challenging in mammalian systems that have 3 genes ( NRXN1-3 ) that encode 2-3 major isoforms, which undergo extensive alternative splicing and generate thousands of transcripts. In contrast to mammals, C. elegans has a single gene, nrx-1 , encoding only long α and short γ isoforms. Neurexins canonically regulate synapse morphology and function in a neuron- and context-specific manner, through mechanisms related to release of chemical neurotransmitters and receptors. Whether neurexins ( nrx-1 ) impact other secretory molecules such as neuropeptides (NPs) and NP containing dense-core vesicles (DCVs) is not well understood. Here, we report that loss of nrx-1 increases the release of multiple NPs from cholinergic motor neurons in C. elegans . Using tissue specific expression and degradation of endogenous NRX-1, we find that nrx-1 functions in NP release from cholinergic neurons in a cell-autonomous manner. We confirm that loss of nrx-1 impacts cholinergic active-zone number, but also find it regulates the clustering, distribution, and expression of the DCV protein, IDA-1 (PTPRN), and the DCV secretion regulator, UNC-31 (CADPS). We find that nrx-1 functions to maintain separation and juxtaposition of neurotransmitter and NP release sites and DCV localization. Loss of cholinergic excitation ( unc-17 ) or GABAergic inhibition ( unc-25 ) did not impact cholinergic NP release, but that the increased NP release upon loss of nrx-1 is dependent on the calcium channel unc-2 . We find that neurexins can regulate NP signaling, a novel mechanism to modify circuits and behaviors, and of potential importance for NRXN1 associated human conditions.