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The intestinal epithelial lining rapidly renews every 4 to 5 days. The renewal is facilitated by intestinal stem cells (ISCs) located at the base of the crypt– a gland located at the bottom of each villus. ISCs divide asymmetrically to form new stem cells and progenitor daughter cells. The daughter cells are called transit-amplifying (TA) cells which move upwards along the crypt and either differentiate into absorptive cells– the enterocytes or secretory cells– including the...
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Immunostaining to Visualize Murine Enteric Nervous System Development
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Intestinal smooth muscle phenotype determines enteric neuronal survival via GDNF expression.

T Y Han1, S Lourenssen1, K G Miller1

  • 1Gastrointestinal Diseases Research Unit, Department of Medicine, Queen's University, Kingston, Ontario, Canada.

Neuroscience
|February 7, 2015
PubMed
Summary

Intestinal smooth muscle cell proliferation supports the enteric nervous system (ENS) by releasing glial cell-derived neurotrophic factor (GDNF). Chronic inflammation may impair this mechanism, leading to neural damage and strictures.

Keywords:
GDNFintestineneuronneurotrophinsmooth muscle proliferation

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Area of Science:

  • Gastroenterology
  • Neuroscience
  • Cell Biology

Background:

  • Intestinal inflammation causes neuronal damage and smooth muscle cell proliferation.
  • Glial cell-derived neurotrophic factor (GDNF) is crucial for enteric nervous system (ENS) development and survival.
  • Inflammatory cytokines upregulate GDNF in intestinal smooth muscle cells (ISMC).

Purpose of the Study:

  • To investigate the relationship between ISMC proliferation and GDNF expression.
  • To determine the role of GDNF in supporting neuronal survival and re-innervation in the inflamed intestine.

Main Methods:

  • Co-culture of myenteric neurons and ISMC.
  • Polymerase chain reaction (PCR) and western blotting to assess GDNF expression and CSMC proliferation.
  • Immunocytochemistry in a TNBS-induced colitis model.
  • Inhibition of GDNF signaling using vandetanib and siGDNF.

Main Results:

  • GDNF and fetal calf serum (FCS) supported neuronal survival in co-cultures.
  • GDNF, but not FCS, supported neuronal survival in low-density cultures lacking ISMC contact.
  • Early-passage colonic circular smooth muscle cells (CSMC) proliferation correlated with GDNF expression and supported neuronal survival.
  • GDNF was selectively expressed in proliferating CSMC in TNBS-induced colitis.
  • High-passage CSMC showed reduced GDNF expression and failed to support neuronal survival.

Conclusions:

  • Smooth muscle proliferation in the inflamed intestine supports the ENS and its re-innervation via GDNF expression.
  • A compromised smooth muscle phenotype in chronic inflammation may result in progressive neural damage.
  • Failure of this homeostatic mechanism could lead to intestinal stricture formation in conditions like inflammatory bowel disease (IBD).