Mutations in the Neuroligin-3 (
Nlgn3) gene are implicated in autism spectrum disorder (ASD) and gastrointestinal (GI) dysfunction, but cellular
Nlgn3 expression in the enteric nervous system remains to be characterised. We combined RNAScope in situ hybridization and immunofluorescence to measure
Nlgn3
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Mutations in the Neuroligin-3 (
Nlgn3) gene are implicated in autism spectrum disorder (ASD) and gastrointestinal (GI) dysfunction, but cellular
Nlgn3 expression in the enteric nervous system remains to be characterised. We combined RNAScope in situ hybridization and immunofluorescence to measure
Nlgn3 mRNA expression in cholinergic and VIP-expressing submucosal neurons, nitrergic and calretinin-containing myenteric neurons and glial cells in both WT and
Nlgn3R451C mutant mice. We measured
Nlgn3 mRNA neuronal and glial expression via quantitative three-dimensional image analysis. To validate dual RNAScope/immunofluorescence data, we interrogated available single-cell RNA sequencing (scRNASeq) data to assess for
Nlgn3,
Nlgn1,
Nlgn2 and their binding partners,
Nrxn1-3,
MGDA1 and
MGDA2, in enteric neural subsets. Most submucosal and myenteric neurons expressed
Nlgn3 mRNA. In contrast to other
Nlgns and binding partners,
Nlgn3 was strongly expressed in enteric glia, suggesting a role for neuroligin-3 in mediating enteric neuron–glia interactions. The autism-associated R451C mutation reduces
Nlgn3 mRNA expression in cholinergic but not in VIPergic submucosal neurons. In the myenteric plexus,
Nlgn3 mRNA levels are reduced in calretinin, nNOS-labelled neurons and S100 β -labelled glia. We provide a comprehensive cellular profile for neuroligin-3 expression in ileal neuronal subpopulations of mice expressing the R451C autism-associated mutation in
Nlgn3, which may contribute to the understanding of the pathophysiology of GI dysfunction in ASD.
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