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Related Experiment Video

Updated: Apr 12, 2026

Immunostaining to Visualize Murine Enteric Nervous System Development
07:54

Immunostaining to Visualize Murine Enteric Nervous System Development

Published on: April 29, 2015

12.0K

Immunostaining to visualize murine enteric nervous system development.

Amanda J Barlow-Anacker1, Christopher S Erickson1, Miles L Epstein2

  • 1Department of Surgery, University of Wisconsin School of Medicine and Public Health.

Journal of Visualized Experiments : Jove
|May 21, 2015
PubMed
Summary
This summary is machine-generated.

This study introduces a reliable method for visualizing cholinergic neurons in the embryonic enteric nervous system using a specific antibody. This technique aids in understanding neural crest cell differentiation and neurotransmitter expression during gut development.

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Related Experiment Videos

Last Updated: Apr 12, 2026

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Area of Science:

  • Neuroscience
  • Developmental Biology
  • Gastroenterology

Background:

  • The enteric nervous system (ENS) originates from neural crest cells that migrate and differentiate within the developing gut.
  • Cholinergic neurons, crucial for ENS function, are identified by choline acetyltransferase (ChAT) expression.
  • Previous methods for visualizing ChAT in the peripheral nervous system were limited by antibody specificity.

Purpose of the Study:

  • To establish a reliable protocol for visualizing cholinergic neurons in the embryonic murine gastrointestinal tract.
  • To address limitations of existing antibodies for detecting peripheral ChAT during embryogenesis.
  • To compare antibody-based detection with genetic reporters for ChAT reliability.

Main Methods:

  • Dissection of murine embryonic gastrointestinal tracts.
  • Fixation and immunostaining protocols optimized for embryonic tissues.
  • Utilizing an antibody against placental choline acetyltransferase (ChAT) for broad specificity.

Main Results:

  • The placental ChAT antibody successfully visualized embryonic enteric cholinergic neurons.
  • This antibody demonstrated greater reliability compared to genetic reporters during embryogenesis.
  • The protocol enables detailed visualization of ENS neurotransmitter expression in developing mouse embryos.

Conclusions:

  • A robust immunostaining technique using a placental ChAT antibody is presented for studying the embryonic ENS.
  • This method overcomes previous limitations in visualizing peripheral cholinergic neurons.
  • The protocol facilitates research into neural crest cell differentiation and ENS development.