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Robust, 3-Dimensional Visualization of Human Colon Enteric Nervous System Without Tissue Sectioning.

Kahleb D Graham1, Silvia Huerta López2, Rajarshi Sengupta3

  • 1Children's Hospital of Philadelphia Research Institute, Abramson Research Center, Philadelphia, Pennsylvania; Cincinnati Children's Hospital Medical Center and the Department of Pediatrics at University of Cincinnati College of Medicine, Cincinnati, Ohio.

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Summary
This summary is machine-generated.

A new 3D imaging technique reveals detailed human enteric nervous system (ENS) structure, improving understanding of bowel innervation and motility disorders.

Keywords:
AcetylcholineHirschsprung DiseaseMyenteric PlexusNitric OxideSubmucosal Ganglia

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Area of Science:

  • Gastroenterology
  • Neuroscience
  • Pathology

Background:

  • Traditional 2D pathology sections offer limited insight into the complex 3D structure of the human enteric nervous system (ENS).
  • Accurate analysis of bowel innervation is crucial for diagnosing motility disorders.

Purpose of the Study:

  • To develop and validate a novel 3D imaging technique for visualizing the human ENS and other intramural cells in full-thickness colon tissue.
  • To enable quantitative analysis of ENS components without traditional tissue sectioning.

Main Methods:

  • A method combining tissue clearing, immunohistochemistry, confocal microscopy, and 3D quantitative analysis was applied to mouse and human colon tissues.
  • Analysis of 280 adult human colon Z-stacks and tissues from Hirschsprung disease patients was performed.
  • A panel of 16 antibodies was validated for imaging neurons, glia, interstitial cells of Cajal, and macrophages.

Main Results:

  • The 3D imaging method successfully visualized the ENS and other intramural cells in full-thickness human colon.
  • Quantitative analysis revealed myenteric plexus density (24.5% ± 2.4% of area) and ganglion size (3,527,678 ± 573,832 mm³).
  • Neuron subtype analysis showed 56% of myenteric neurons stained positive for neuronal nitric oxide synthase and 33% produce acetylcholine; variability in ENS density across millimeters was observed.

Conclusions:

  • The developed 3D imaging technique provides significantly more information on ENS structure compared to conventional 2D sectioning.
  • This advanced imaging approach holds potential for improved diagnosis of human bowel motility disorders and other gastrointestinal diseases.