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Updated: Jul 19, 2026
07:54
Immunostaining to Visualize Murine Enteric Nervous System Development
Published on: April 29, 2015
High affinity choline transporter immunoreactivity in rat ileum myenteric nerves
Andrea M Harrington1, J M Hutson, Bridget R Southwell
1Gut Motility Laboratory, Murdoch Children's Research Institute, Royal Children's Hospital, Parkville, Australia.
Cell and Tissue Research
|November 10, 2006
Summary
An antibody to the choline transporter (CHT) labels nerve fibers and neurons in the rat ileum's enteric nervous system (ENS). While CHT immunoreactivity colocalizes with vesicular acetylcholine transporter (VAChT), it does not label all cholinergic neurons.
Area of Science:
- Neuroscience
- Gastroenterology
- Immunohistochemistry
Background:
- The choline transporter (CHT) is crucial for acetylcholine (ACh) uptake in the central nervous system.
- CHT's role in the enteric nervous system (ENS) remains largely unexamined.
- Previous studies focused on CHT in the central nervous system, leaving the ENS unexplored.
Purpose of the Study:
- To investigate the localization of CHT immunoreactivity (CHT-IR) in the rat ileum's ENS.
- To determine the colocalization of CHT-IR with markers for cholinergic, tachykinergic, and nitrergic neurons.
- To assess the utility of anti-CHT antibodies for labeling ENS components.
Main Methods:
- Rat ileum segments were processed for whole-mounts and cryosections.
- Immunohistochemistry was performed using an anti-CHT antiserum.
- Double-labeling with antibodies against nitric oxide synthase, substance P (SP), choline acetyltransferase (cChAT), and vesicular acetylcholine transporter (VAChT) was employed.
Main Results:
- CHT-IR was detected in nerve fibers and myenteric ganglia of the rat ileum.
- CHT-IR colocalized with VAChT-IR in the myenteric plexus and circular muscle, but not universally.
- No CHT-IR neurons were found to be cChAT-immunoreactive; however, some CHT-IR neurons and fibers showed SP immunoreactivity.
Conclusions:
- Anti-CHT antiserum successfully labels neurons and nerve fibers within the rat ENS.
- CHT-IR in the ENS does not exclusively identify cholinergic neurons marked by cChAT but does identify VAChT-immunoreactive nerve fibers.
- The findings suggest CHT is a marker for a subset of ENS neurons and nerve fibers, including some not expressing VAChT.
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