Skip to main content

Advertisement

Log in

Altered differentiation of enteric neural crest-derived cells from endothelin receptor-B null mouse model of Hirschsprung’s disease

  • Original Article
  • Published:
Pediatric Surgery International Aims and scope Submit manuscript

Abstract

Purpose

Hirschsprung’s disease (HD) is caused by a failure of enteric neural crest-derived cells (ENCC) to colonize the bowel, resulting in an absence of the enteric nervous system (ENS). Previously, we developed a Sox10 transgenic version of the Endothelin receptor-B (Ednrb) mouse to visualize ENCC with the green fluorescent protein, Venus. The aim of this study was to isolate Sox10-Venus+ cells, which are differentiated neurons and glial cells in the ENS, and analyze these cells using Sox10-Venus mice gut.

Methods

The mid-and hindgut of Sox10-Venus+/Ednrb +/+ and Sox10-Venus+/Ednrb / at E13.5 and E15.5 were dissected and cells were dissociated. Sox10-Venus+ cells were then isolated. Expression of PGP9.5 and GFAP were evaluated neurospheres using laser scanning microscopy.

Results

7 days after incubation, Sox10-Venus+ cells colonized the neurosphere. There were no significant differences in PGP9.5 expressions on E13.5 and E15.5. GFAP was significantly increased in HD compared to controls on E15.5 (P < 0.05).

Conclusions

Our results suggest increased glial differentiation causes an imbalance in ENCC lineages, leading to a disruption of normal ENS development in this HD model. Isolation of ENCC provides an opportunity to investigate the ENS with purity and might be a useful tool for modeling cell therapy approaches to HD.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Goldstein AM, Hofstra RMW, Burns AJ (2013) Building a brain in the gut: development of the enteric nervous system. Clin Genet 83:307–316

    Article  CAS  PubMed  Google Scholar 

  2. Xie D, Croaker GDH, Li JM, Song ZM (2016) Reduced cell proliferation and increased apoptosis in the hippocampal formation in a rat model of Hirschsprung’s disease. Brain Res 1642:79–86

    Article  CAS  PubMed  Google Scholar 

  3. Musser MA, Southard-Smith EM (2013) Balancing on the crest—evidence for disruption of the enteric ganglia via inappropriate lineage segregation and consequences for gastrointestinal function. Dev Biol 382:356–364

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Wegner M (1999) From head to toes: the multiple facets of Sox proteins. Nucl. Acids Res 27(6):1409–1420

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Kawaguchi J, Nichols J, Gierl MS et al (2010) Isolation and propagation of enteric neural crest progenitor cells from mouse embryonic stem cells and embryos. Development 137:693–704

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Hearn CJ, Murphy M, Newgreen D (1998) GDNF and ET-3 differentially modulate the numbers of avian enteric neural crest cells and enteric neurons in vitro. Dev Biol 197:93–105

    Article  CAS  PubMed  Google Scholar 

  7. Nagy N, Goldstein AM (2006) Endothelin-3 regulated neural crest cell proliferation and differentiation in the hindgut enteric nervous system. Dev Biol 293:203–217

    Article  CAS  PubMed  Google Scholar 

  8. Wu JJ, Chen JX, Rothman TP et al (1999) Inhibition of in vitro enteric neuronal development by endothelin-3: mediation by endothelin B receptors. Development 126:1161–1173

    CAS  PubMed  Google Scholar 

  9. Barlow A, de Graaff E, Pachnis V (2003) Enteric nervous system progenitors are coordinately controlled by the G protein-coupled receptor EDNRB and the receptor tyrosine kinase RET. Neuron 40:905–916

    Article  CAS  PubMed  Google Scholar 

  10. Miyahara K, Kato Y, Koga H et al (2010) Abnormal enteric innervation identified without histopathologic staining in aganglionic colorectum from a mouse model of Hirschsprung’s disease. J Pediatr Surg 45:2403–2407

    Article  PubMed  Google Scholar 

  11. Shibata S, Yasuda A, Renault-Mihara F et al (2010) Sox10-Venus mice: a new tool for real-time labeling of neural crest lineage cells and oligodendrocytes. Mol Brain 3:31

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Bondurand N, Natarajan D, Thapar N et al (2003) Neuron and glia generating progenitors of the mammalian enteric nervous system isolated from fortal and postnatal gut cultures. Developement 130:6387–6400

    CAS  Google Scholar 

  13. Grundmann D, Klotz M, Rabe H et al (2015) Isolation of high-purity myenteic plexus from adult human and mouse gastrointestinal tract. Sci Rep 20(5):9226

    Article  CAS  Google Scholar 

  14. Xu Q, Heanue T, Pachnis V (2014) Travelling within the fetal gut: simple rules for an arduous journey. BMC Bio 30(12):50

    Article  CAS  Google Scholar 

  15. Amiel J, Sproat-Emison E, Garcia-Barcelo M et al (2008) Hirschsprung disease, associated syndromes and genetics: a review. J Med Genet 45(1):1–14

    Article  CAS  PubMed  Google Scholar 

  16. Lake JL, Heuckroth RO (2013) Enteric nervous system development: migration, differentiation, and disease. Am J Physiol Gastrointest Liver Physiol 305(1):G1–24

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Young HM, Bergner AJ, Muller T (1993) Acquisition of neuronal and glial markers by neural crest-derived cells in the mouse intestine. J Comp Neurol 456(1):1–11

    Article  Google Scholar 

  18. Heanue TA, Pachnis V (2007) Enteric nervous system development and Hirschsprung’s disease: advances in genetic and stem cell studies. Nat Rev Neurosci 8(6):466–479

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank Dr. Hiroshi Koide for excellent technical support and their critical advice.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Naho Fujiwara.

Ethics declarations

Funding

This work is supported by JSPS KAKENHI Grant Number 16K20347.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Fujiwara, N., Miyahara, K., Nakazawa-Tanaka, N. et al. Altered differentiation of enteric neural crest-derived cells from endothelin receptor-B null mouse model of Hirschsprung’s disease. Pediatr Surg Int 32, 1095–1101 (2016). https://doi.org/10.1007/s00383-016-3964-4

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00383-016-3964-4

Keywords

Navigation