Genome-wide analysis reveals pathways important for the development and maturation of excitatory synaptic connections to GABAergic neurons

Abstract

A high degree of cell and circuit-specific regulation has presented challenges for efforts to precisely define molecular mechanisms controlling synapse formation and maturation. Here, we pursue an unbiased forward genetic approach to identify C. elegans genes involved in the formation and maturation of cholinergic synaptic connections with GABAergic motor neurons as indicated by the distribution of GFP-tagged postsynaptic acetylcholine receptors (AChR) on GABAergic dendrites. We identified mutations in 3 genes that identify key processes in synapse/circuit maturation. Mutation of the RUN domain (RPIP8, UNC-14, and NESCA) cargo adaptor gene unc-14 dramatically impacts overall GABAergic neuron morphology and dendritic spines. Mutation of the nicotinic acetylcholine alpha subunit gene unc-63 causes a failure in AChR assembly in GABAergic neurons but does not significantly alter dendritic spine structure or abundance. Finally, a mutation in the Liprin- synaptic scaffold gene syd-2 severely disrupts both dendritic spines and AChR localization. The identification of these three genes from our screen highlights how mechanisms for cargo trafficking, receptor assembly, and synapse structural organization each make distinct contributions to synapse assembly and circuit connectivity.