Quantitative Spatial Analysis of Neuroligin-3 mRNA Expression in the Enteric Nervous System Reveals a Potential Role in Neuronal–Glial Synapses and Reduced Expression in <i>Nlgn3<sup>R451C</sup></i> Mice
Mutations in the Neuroligin-3 (<i>Nlgn3</i>) gene are implicated in autism spectrum disorder (ASD) and gastrointestinal (GI) dysfunction, but cellular <i>Nlgn3</i> expression in the enteric nervous system remains to be characterised. We combined RNAScope in situ hybridization...
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MDPI AG
2023-06-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/13/7/1063 |
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| author | Madushani Herath Ellie Cho Ulrika Marklund Ashley E. Franks Joel C. Bornstein Elisa L. Hill-Yardin |
| author_facet | Madushani Herath Ellie Cho Ulrika Marklund Ashley E. Franks Joel C. Bornstein Elisa L. Hill-Yardin |
| author_sort | Madushani Herath |
| collection | DOAJ |
| description | Mutations in the Neuroligin-3 (<i>Nlgn3</i>) gene are implicated in autism spectrum disorder (ASD) and gastrointestinal (GI) dysfunction, but cellular <i>Nlgn3</i> expression in the enteric nervous system remains to be characterised. We combined RNAScope in situ hybridization and immunofluorescence to measure <i>Nlgn3</i> mRNA expression in cholinergic and VIP-expressing submucosal neurons, nitrergic and calretinin-containing myenteric neurons and glial cells in both WT and <i>Nlgn3<sup>R451C</sup></i> mutant mice. We measured <i>Nlgn3</i> 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 <i>Nlgn3</i>, <i>Nlgn1</i>, <i>Nlgn2</i> and their binding partners, <i>Nrxn1-3</i>, <i>MGDA1</i> and <i>MGDA2</i>, in enteric neural subsets. Most submucosal and myenteric neurons expressed <i>Nlgn3</i> mRNA. In contrast to other <i>Nlgns</i> and binding partners, <i>Nlgn3</i> was strongly expressed in enteric glia, suggesting a role for neuroligin-3 in mediating enteric neuron–glia interactions. The autism-associated R451C mutation reduces <i>Nlgn3</i> mRNA expression in cholinergic but not in VIPergic submucosal neurons. In the myenteric plexus, <i>Nlgn3</i> 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 <i>Nlgn3,</i> which may contribute to the understanding of the pathophysiology of GI dysfunction in ASD. |
| first_indexed | 2024-03-11T01:15:46Z |
| format | Article |
| id | doaj.art-8d652b5e6c184db0b5312a586c85ac07 |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-11T01:15:46Z |
| publishDate | 2023-06-01 |
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| series | Biomolecules |
| spelling | doaj.art-8d652b5e6c184db0b5312a586c85ac072023-11-18T18:30:53ZengMDPI AGBiomolecules2218-273X2023-06-01137106310.3390/biom13071063Quantitative Spatial Analysis of Neuroligin-3 mRNA Expression in the Enteric Nervous System Reveals a Potential Role in Neuronal–Glial Synapses and Reduced Expression in <i>Nlgn3<sup>R451C</sup></i> MiceMadushani Herath0Ellie Cho1Ulrika Marklund2Ashley E. Franks3Joel C. Bornstein4Elisa L. Hill-Yardin5Department of Anatomy & Physiology, University of Melbourne, Parkville, VIC 3010, AustraliaBiological Optical Microscopy Platform, University of Melbourne, Parkville, VIC 3010, AustraliaDivision of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, 171 77 Stockholm, SwedenDepartment of Microbiology, Anatomy Physiology & Pharmacology, School of Agriculture, Biomedicine and Environment, La Trobe University, Bundoora, VIC 3086, AustraliaDepartment of Anatomy & Physiology, University of Melbourne, Parkville, VIC 3010, AustraliaSchool of Health and Biomedical Sciences, STEM College, RMIT University, Bundoora, VIC 3083, AustraliaMutations in the Neuroligin-3 (<i>Nlgn3</i>) gene are implicated in autism spectrum disorder (ASD) and gastrointestinal (GI) dysfunction, but cellular <i>Nlgn3</i> expression in the enteric nervous system remains to be characterised. We combined RNAScope in situ hybridization and immunofluorescence to measure <i>Nlgn3</i> mRNA expression in cholinergic and VIP-expressing submucosal neurons, nitrergic and calretinin-containing myenteric neurons and glial cells in both WT and <i>Nlgn3<sup>R451C</sup></i> mutant mice. We measured <i>Nlgn3</i> 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 <i>Nlgn3</i>, <i>Nlgn1</i>, <i>Nlgn2</i> and their binding partners, <i>Nrxn1-3</i>, <i>MGDA1</i> and <i>MGDA2</i>, in enteric neural subsets. Most submucosal and myenteric neurons expressed <i>Nlgn3</i> mRNA. In contrast to other <i>Nlgns</i> and binding partners, <i>Nlgn3</i> was strongly expressed in enteric glia, suggesting a role for neuroligin-3 in mediating enteric neuron–glia interactions. The autism-associated R451C mutation reduces <i>Nlgn3</i> mRNA expression in cholinergic but not in VIPergic submucosal neurons. In the myenteric plexus, <i>Nlgn3</i> 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 <i>Nlgn3,</i> which may contribute to the understanding of the pathophysiology of GI dysfunction in ASD.https://www.mdpi.com/2218-273X/13/7/1063neuroligin-3autismenteric nervous systemgliaileumRNAScope |
| spellingShingle | Madushani Herath Ellie Cho Ulrika Marklund Ashley E. Franks Joel C. Bornstein Elisa L. Hill-Yardin Quantitative Spatial Analysis of Neuroligin-3 mRNA Expression in the Enteric Nervous System Reveals a Potential Role in Neuronal–Glial Synapses and Reduced Expression in <i>Nlgn3<sup>R451C</sup></i> Mice Biomolecules neuroligin-3 autism enteric nervous system glia ileum RNAScope |
| title | Quantitative Spatial Analysis of Neuroligin-3 mRNA Expression in the Enteric Nervous System Reveals a Potential Role in Neuronal–Glial Synapses and Reduced Expression in <i>Nlgn3<sup>R451C</sup></i> Mice |
| title_full | Quantitative Spatial Analysis of Neuroligin-3 mRNA Expression in the Enteric Nervous System Reveals a Potential Role in Neuronal–Glial Synapses and Reduced Expression in <i>Nlgn3<sup>R451C</sup></i> Mice |
| title_fullStr | Quantitative Spatial Analysis of Neuroligin-3 mRNA Expression in the Enteric Nervous System Reveals a Potential Role in Neuronal–Glial Synapses and Reduced Expression in <i>Nlgn3<sup>R451C</sup></i> Mice |
| title_full_unstemmed | Quantitative Spatial Analysis of Neuroligin-3 mRNA Expression in the Enteric Nervous System Reveals a Potential Role in Neuronal–Glial Synapses and Reduced Expression in <i>Nlgn3<sup>R451C</sup></i> Mice |
| title_short | Quantitative Spatial Analysis of Neuroligin-3 mRNA Expression in the Enteric Nervous System Reveals a Potential Role in Neuronal–Glial Synapses and Reduced Expression in <i>Nlgn3<sup>R451C</sup></i> Mice |
| title_sort | quantitative spatial analysis of neuroligin 3 mrna expression in the enteric nervous system reveals a potential role in neuronal glial synapses and reduced expression in i nlgn3 sup r451c sup i mice |
| topic | neuroligin-3 autism enteric nervous system glia ileum RNAScope |
| url | https://www.mdpi.com/2218-273X/13/7/1063 |
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