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Department of Pharmacology

 
Author(s): 
Hockley, J, Taylor, T, Callejo, G, Wilbrey, A, Gutterridge, A, Bach, K, Winchester, W, Bulmer, D, McMurray, G, Smith, ESJ
Abstract: 

Objective Integration of nutritional, microbial and inflammatory events along the gut-brain axis can alter bowel physiology and organism behaviour. Colonic sensory neurones activate reflex pathways and give rise to conscious sensation, but the diversity and division of function within these neurones is poorly understood. The identification of signalling pathways contributing to visceral sensation is constrained by a paucity of molecular markers. Here we address this by comprehensive transcriptomic profiling and unsupervised clustering of individual mouse colonic sensory neurones.

Design Unbiased single-cell RNA-sequencing was performed on retrogradely traced mouse colonic sensory neurones isolated from both thoracolumbar (TL) and lumbosacral (LS) dorsal root ganglia associated with lumbar splanchnic and pelvic spinal pathways, respectively. Identified neuronal subtypes were validated by single-cell qRT-PCR, immunohistochemistry and Ca2+-imaging.

Results Transcriptomic profiling and unsupervised clustering of 314 colonic sensory neurones revealed 7 neuronal subtypes. Of these, 5 neuronal subtypes accounted for 99% of TL neurones, with LS neurones almost exclusively populating the remaining 2 subtypes. We identify and classify neurones based on novel subtype-specific marker genes using single-cell qRT-PCR and immunohistochemistry to validate subtypes derived from RNA-sequencing. Lastly, functional Ca2+-imaging was conducted on colonic sensory neurons to demonstrate subtype-selective differential agonist activation.

Conclusions We identify 7 subtypes of colonic sensory neurones using unbiased single-cell RNA-sequencing and confirm translation of patterning to protein expression, describing sensory diversity encompassing all modalities of colonic neuronal sensitivity. These results provide a pathway to molecular interrogation of colonic sensory innervation in health and disease, together with identifying novel targets for drug development.

Publication ID: 
966079
Published date: 
10 February 2018 (Accepted for publication)
Publication source: 
manual
Publication type: 
Journal articles
Journal name: 
Gut
Publication volume: 
Publisher: 
BMJ
Parent title: 
Edition: 
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