# CONGENITAL MEGACOLON: TISSUE INTERACTIONS IN DEVELOPMENT

> **NIH NIH R01** · COLUMBIA UNIV NEW YORK MORNINGSIDE · 1994 · $215,404

## Abstract

The enteric nervous system is formed by precursors that migrate to the
bowel from the neural crest.  In order to determine why crest-derived
cells stop migrating and begin to differentiate in the gut, we have
studied a mutant mouse, the lethal spotted (ls/ls), in which the terminal
bowel is congenitally aganglionic.  We have shown that the defect in
these animals arises, not because their crest-derived cells are abnormal,
but because neither these, nor any other source of crest-derived cells,
is able to colonize the presumptive aganglionic ls/ls gut.  Within this
tissue, molecular components of basal laminae, including laminin, are
overabundant and/or maldistributed.  We have proposed that crest-derived
cells, when they enter the bowel, acquire a nerve-related 110 kDa laminin
receptor which, when activated, induces these cells to withdraw from the
cell cycle, extend processes and cease migrating.  The 110 kDa laminin
binding site is not found on pre- or early migrating crest cells.  We
thus wish to test the hypothesis that the ls/ls defect is due to the
abnormal extracellular matrix (ECM) of the presumptive aganglionic zone.
We propose that migrating crest-derived cells stop, cease to proliferate,
and extend processes prematurely when they encounter this ECM.  As a
result, the distal bowel does not become colonized by crest-derived
precursors of neurons and glia, and there is no accumulation of neurons
at the periphery of the abnormal zone.  We will now test this hypothesis
by using a purified population of crest-derived cells isolated from
within the gut by magnetic immunoselection (employing antibodies to cell
surface molecules expressed in the gut only by crest-derived cells).  The
effects of laminin and a variety of other defined substrates on the
behavior of these cells in vitro will be ascertained.  Parameters to be
studied will include adhesion, migration, proliferation, neurite
extension, and phenotypic expression.  In addition to observation,
quantitative measurements will be made of neural and glial expression,
using immunocytochemical markers.  Immunoselected enteric crest-derived
cells will also be cultured on cyostat sections of control and presumptive
aganglionic ls/ls bowel, in order to determine whether laminin or other
molecules mimic the action of the abnormal ECM of the ls/ls gut on each
the measured parameters of cell behavior.  The effects on these parameters
of synthetic peptides, which encompass neurite-promoting domains of
laminin, antibodies to these domains, and antibodies to the 110 kDa
protein will then be tested on immunoselected crest-derived cells
cultured on laminin or cryostat sections of presumptive aganglionic ls/ls
and control gut.  Finally, the behavior of crest-derived cells and the
effects of potential laminin antagonists will be analyzed by direct
visualization using confocal fluorescence microscopy to observe the cells
as they migrate in vitro within explants of fetal bowel from ls/ls and
control mice.

## Key facts

- **NIH application ID:** 2197523
- **Project number:** 5R01HD017736-10
- **Recipient organization:** COLUMBIA UNIV NEW YORK MORNINGSIDE
- **SAM.gov UEI:** F4N1QNPB95M4
- **Principal Investigator:** VIRGINIA M TENNYSON
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1994
- **Award amount:** $215,404
- **Award type:** 5
- **Project period:** 1983-12-01 → 1997-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/2197523

## Citation

> US National Institutes of Health, RePORTER application 2197523, CONGENITAL MEGACOLON: TISSUE INTERACTIONS IN DEVELOPMENT (5R01HD017736-10). Retrieved via MORVS 2026-10-26 from https://api.morvs.ai/grant/nih/2197523. Licensed CC0.

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