Gastroenterology

Gastroenterology

Volume 130, Issue 4, April 2006, Pages 1221-1232
Gastroenterology

Basic–alimentary tract
The Cell Adhesion Molecule L1 Is Required for Chain Migration of Neural Crest Cells in the Developing Mouse Gut

https://doi.org/10.1053/j.gastro.2006.01.002Get rights and content

Background & Aims: During development, the enteric nervous system is derived from neural crest cells that emigrate from the hindbrain, enter the foregut, and colonize the gut. Defects in neural crest migration can result in intestinal aganglionosis. Hirschsprung’s disease (congenital aganglionosis) is a human condition in which enteric neurons are absent from the distal bowel. A number of clinical studies have implicated the cell adhesion molecule L1 in Hirschsprung’s disease. We examined the role of L1 in the migration of neural crest cells through the developing mouse gut. Methods: A variety of in vitro and in vivo assays were used to examine: (1) the effect of L1 blocking antibodies or exogenous soluble L1 protein known to compromise L1 function on the rate of crest cell migration, (2) the effect of blocking L1 activity on the dynamic behavior of crest cells using time-lapse microscopy, and (3) whether the colonization of the gut by crest cells in L1-deficient mice differs from control mice. Results: We show that L1 is expressed by neural crest cells as they colonize the gut. Perturbation studies show that disrupting L1 activity retards neural crest migration and increases the number of solitary neural crest cells. L1-deficient mice show a small but significant reduction in neural crest cell migration at early developmental stages, but the entire gastrointestinal tract is colonized. Conclusions: L1 is important for the migration of neural crest cells through the developing gut and is likely to be involved in the etiology of Hirschsprung’s disease.

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Animals

In this study, the following mice were used: (1) C57BL/6; (2) RetTGM mice in which complementary DNA encoding tau-EGFP-myc (TGM) had been inserted into the first coding exon of the receptor tyrosine kinase gene, Ret26; (3) DβH-nlacZ mice in which the expression of nlacZ is driven by the human DβH promoter27; and (4) L1-deficient mice.28 The RetTGM and DβH-nlacZ mice were on a C57BL/6 background, and the L1-deficient mice were on a mixed 129 × NMRI background. The genotypes of RetTGM, DβH-nlacZ,

L1 Is Expressed by Enteric Neural Crest Cells

Vagal neural crest cells colonize the embryonic mouse gut in a rostral-to-caudal wave between E10 and E14.5.32 A previous study had shown that cells expressing L1, which were in the correct location to be enteric neuron precursors, were present in the gut of E14.5 mice.33 Double-label immunohistochemistry was performed on whole mounts of E10.5–E14.5 gut using antibodies against L1 and Phox2b. Phox2b has previously been shown to be expressed by all neural crest cells within the embryonic gut.34

Discussion

Some humans with mutations in the L1 gene also have HSCR,13, 14, 15 suggesting that L1 may play a role in the migration of neural crest cells along the human gut. In this study, we used in vitro and in vivo approaches to investigate the role of L1 signaling in enteric neural crest cell migration in mice. We provide the first evidence that L1 is expressed exclusively by neural crest–derived cells in the embryonic mouse gut and is required for their correct migration. Previous studies have shown

References (62)

  • A. Chalazonitis et al.

    Age-dependent differences in the effects of GDNF and NT-3 on the development of neurons and glia from neural crest-derived precursors immunoselected from the fetal rat gutexpression of GFRalpha-1 in vitro and in vivo

    Dev Biol

    (1998)
  • H.M. Young et al.

    A single rostrocaudal colonization of the rodent intestine by enteric neuron precursors is revealed by the expression of Phox2b, Ret, and p75 and by explants grown under the kidney capsule or in organ culture

    Dev Biol

    (1998)
  • H. Ikawa et al.

    Impaired expression of neural cell adhesion molecule L1 in the extrinsic nerve fibers in Hirschsprung’s disease

    J Pediatr Surg

    (1997)
  • M.A. Parisi et al.

    Genetic background modifies intestinal pseudo-obstruction and the expression of a reporter gene in Hox11L1-/- mice

    Gastroenterology

    (2003)
  • R.P. Kapur et al.

    Abnormal microenvironmental signals underlie intestinal aganglionosis in dominant megacolon mutant mice

    Dev Biol

    (1996)
  • A. Auricchio et al.

    Double heterozygosity for a RET substitution interfering with splicing and an EDNRB missense mutation in Hirschsprung disease

    Am J Hum Genet

    (1999)
  • A. Barlow et al.

    Enteric nervous system progenitors are coordinately controlled by the g protein-coupled receptor EDNRB and the receptor tyrosine kinase RET

    Neuron

    (2003)
  • C.L. Yntema et al.

    The origin of intrinsic ganglia of trunk viscera from vagal neural crest in the chick embryo

    J Comp Neurol

    (1954)
  • N.M. Le Douarin et al.

    The migration of neural crest cells to the wall of the digestive tract in avian embryo

    J Embryol Exp Morphol

    (1973)
  • P. Puri et al.

    Experience with Swenson’s operation

  • W.M. Belknap

    The pathogenesis of Hirschsprung disease

    Curr Opin Gastroenterol

    (2002)
  • C.E. Gariepy

    Developmental disorders of the enteric nervous systemgenetic and molecular bases

    J Pediatr Gastroenterol Nutr

    (2004)
  • M.A. Parisi et al.

    Genetics of Hirschsprung disease

    Curr Opin Pediatr

    (2000)
  • J.A. Badner et al.

    A genetic study of Hirschsprung disease

    Am J Hum Genet

    (1990)
  • S. Bolk et al.

    A human model for multigenic inheritancephenotypic expression in Hirschsprung disease requires both the RET gene and a new 9q31 locus

    Proc Natl Acad Sci U S A

    (2000)
  • M.M. Carrasquillo et al.

    Genome-wide association study and mouse model identify interaction between RET and EDNRB pathways in Hirschsprung disease

    Nat Genet

    (2002)
  • A.S. McCallion et al.

    Phenotype variation in two-locus mouse models of Hirschsprung diseasetissue-specific interaction between Ret and Ednrb

    Proc Natl Acad Sci U S A

    (2003)
  • V.A. Cantrell et al.

    Interactions between Sox10 and EdnrB modulate penetrance and severity of aganglionosis in the Sox10Dom mouse model of Hirschsprung disease

    Hum Mol Genet

    (2004)
  • S. Kenwrick et al.

    Neural cell recognition molecule L1relating biological complexity to human disease mutations

    Hum Mol Genet

    (2000)
  • N. Okamoto et al.

    Hydrocephalus and Hirschsprung’s disease in a patient with a mutation of L1CAM

    J Med Genet

    (1997)
  • R.M. Hofstra et al.

    Hirschsprung disease and L1CAMis the disturbed sex ratio caused by L1CAM mutations?

    J Med Genet

    (2002)

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    Supported by the National Health and Medical Research Council (project grant 350311 to R.B.A. and H.M.Y., Senior Research Fellowship 170224 to H.M.Y., and C. J. Martin Fellowship 007144 to R.B.A.), ANZ Trust (to R.B.A.), the University of Melbourne Early Career Research Grant (to R.B.A.), and the Sir Colin and Lady MacKenzie Trust Award (to R.B.A.).

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    Copyright © 2006 American Gastroenterological Association Institute. Published by Elsevier Inc. All rights reserved.