Abstract
The gut-brain axis has received increasing attention recently due to evidence that colonic microbes can affect brain function and behavior. However, little is known about the innervation of the colon by a major component of the gut-brain axis, vagal afferent neurons. Furthermore, it is currently unknown whether individual NG neurons or DRG neurons innervate both the proximal and distal colon. We aimed to quantify the number of vagal and spinal afferent neurons that innervate the colon; and determine whether these individual neurons simultaneously innervate the mouse proximal and distal colon. C57Bl/6 mice received injections of a combination of retrograde tracers that were either injected into the muscularis externa of the proximal or the distal colon: fast blue, DiI and DiO. Five to seven percent of lumbosacral and thoracolumbar spinal afferent neurons, and 25% of vagal afferent neurons were labelled by injections of DiI and DiO into the colon. We also found that approximately 8% of NG neurons innervate the distal colon. Ten percent of labeled thoracolumbar and 15% of labeled lumbosacral DRG neurons innervate both the distal and proximal colon. Eighteen percent of labeled NG neurons innervated both the distal and proximal colon. In conclusion, vagal afferent innervation of the distal colon is less extensive than the proximal colon, whereas a similar gradient was not observed for the spinal afferent innervation. Furthermore, overlap appears to exist between the receptive fields of vagal and spinal afferent neurons that innervate the proximal and distal colon.
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This research was supported by grants from the Natural Sciences and Engineering Research Council of Canada and the Canadian Institutes of Health Research.
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AL designed the study. AT and SO conducted the retrograde surgeries, tissue sectioning and analysis. AT, SO, DR and AL wrote the paper. All authors approved the final manuscript.
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All experimental protocols were carried out in accordance with the guidelines of the Canadian Council on Animal Care and approved by Queen’s University Animal Care Committee (Protocol #2018-1816). This article does not contain any studies with human participants performed by any of the authors.
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Osman, S., Tashtush, A., Reed, D.E. et al. Analysis of the spinal and vagal afferent innervation of the mouse colon using neuronal retrograde tracers. Cell Tissue Res 392, 659–670 (2023). https://doi.org/10.1007/s00441-023-03769-3
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DOI: https://doi.org/10.1007/s00441-023-03769-3