Gut-derived factors promote neurogenesis of CNS-neural stem cells and nudge their differentiation to an enteric-like neuronal phenotype

Subhash Kulkarni, Bende Zou, Jesse Hanson, Maria Micci, Gunjan Tiwari, Laren Becker, Martin Kaiser, Xinmin Xie, Pankaj Jay Pasricha

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Recent studies have explored the potential of central nervous system-derived neural stem cells (CNSNSC) to repopulate the enteric nervous system. However, the exact phenotypic fate of gut-transplanted CNS-NSC has not been characterized. The aim of this study was to investigate the effect of the gut microenvironment on phenotypic fate of CNS-NSC in vitro. With the use of Transwell culture, differentiation of mouse embryonic CNS-NSC was studied when cocultured without direct contact with mouse intestinal longitudinal muscle-myenteric plexus preparations (LM-MP) compared with control noncocultured cells, in a differentiating medium. Differentiated cells were analyzed by immunocytochemistry and quantitative RT-PCR to assess the expression of specific markers and by whole cell patch-clamp studies for functional characterization of their phenotype. We found that LM-MP cocultured cells had a significant increase in the numbers of cells that were immune reactive against the panneuronal marker β-tubulin, neurotransmitters neuronal nitric oxide synthase (nNOS), choline acetyltransferase (ChAT), and neuropeptide vasoactive intestinal peptide (VIP) and showed an increase in expression of these genes, compared with control cells. Whole cell patch-clamp analysis showed that coculture with LM-MP decreases cell excitability and reduces voltage-gated Na + currents but significantly enhances A-current and late afterhyperpolarization (AHP) and increases the expression of the four AHP-generating Ca2 +-dependent K + channel genes (KCNN), compared with control cells. In a separate experiment, differentiation of LM-MP cocultured CNS-NSC produced a significant increase in the numbers of cells that were immune reactive against the neurotransmitters nNOS, ChAT, and the neuropeptide VIP compared with CNS-NSC differentiated similarly in the presence of neonatal brain tissue. Our results show that the gut microenvironment induces CNS-NSC to produce neurons that share some of the characteristics of classical enteric neurons, further supporting the therapeutic use of these cells for gastrointestinal disorders.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume301
Issue number4
DOIs
StatePublished - Oct 2011

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proctolin
Neural Stem Cells
Neurogenesis
Phenotype
Myenteric Plexus
Muscles
Nitric Oxide Synthase Type I
Choline O-Acetyltransferase
Vasoactive Intestinal Peptide
Neuropeptides
Neurotransmitter Agents
Cell Count
Enteric Nervous System
Neurons
Therapeutic Uses
Tubulin
Coculture Techniques

Keywords

  • Afterhyperpolarization
  • Ca2 -dependent K channel genes
  • Central nervous system
  • Enteric nervous system
  • Longitudinal musclemyenteric plexus
  • Neuronal nitric oxide synthase

ASJC Scopus subject areas

  • Gastroenterology
  • Physiology (medical)
  • Physiology
  • Hepatology

Cite this

Gut-derived factors promote neurogenesis of CNS-neural stem cells and nudge their differentiation to an enteric-like neuronal phenotype. / Kulkarni, Subhash; Zou, Bende; Hanson, Jesse; Micci, Maria; Tiwari, Gunjan; Becker, Laren; Kaiser, Martin; Xie, Xinmin; Pasricha, Pankaj Jay.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 301, No. 4, 10.2011.

Research output: Contribution to journalArticle

Kulkarni, Subhash ; Zou, Bende ; Hanson, Jesse ; Micci, Maria ; Tiwari, Gunjan ; Becker, Laren ; Kaiser, Martin ; Xie, Xinmin ; Pasricha, Pankaj Jay. / Gut-derived factors promote neurogenesis of CNS-neural stem cells and nudge their differentiation to an enteric-like neuronal phenotype. In: American Journal of Physiology - Gastrointestinal and Liver Physiology. 2011 ; Vol. 301, No. 4.
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AU - Kaiser, Martin

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AU - Pasricha, Pankaj Jay

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