Gastroenterology

Gastroenterology

Volume 117, Issue 3, September 1999, Pages 584-594
Gastroenterology

Alimentary Tract
Development of interstitial cells of Cajal and pacemaking in mice lacking enteric nerves,☆☆

https://doi.org/10.1016/S0016-5085(99)70451-8Get rights and content

Abstract

Background & Aims: Development of interstitial cells of Cajal (ICC) requires signaling via Kit receptors. Kit is activated by stem cell factor (SCF), but the source of SCF in the bowel wall is unclear and controversy exists about whether enteric neurons express the SCF required for ICC development. Methods: Glial cell line–derived neurotrophic factor (GDNF) knockout mice, which lack enteric neurons throughout most of the gut, were used to determine whether neurons are necessary for ICC development. ICC distributions were determined with Kit immunofluorescence, and function of ICC was determined by intracellular electrical recording. Results: ICC were normally distributed throughout the gastrointestinal tracts of GDNF−/− mice. Intracellular recordings from aganglionic gastrointestinal muscles showed normal slow wave activity at birth in the stomach and small intestine. Slow waves developed normally in aganglionic segments of small bowel placed into organ culture at birth. Quantitative polymerase chain reaction showed similar expression of SCF in the muscles of animals with and without enteric neurons. Expression of SCF was demonstrated in isolated intestinal smooth muscle cells. Conclusions: These data suggest that enteric neurons are not required for the development of functional ICC. The circular smooth muscle layer, which develops before ICC, may be the source of SCF required for ICC development.

GASTROENTEROLOGY 1999;117:584-594

Section snippets

Animals

GDNF knockout mice (D0) were obtained from the Laboratory of Mammalian Genes and Development at the National Institutes of Child Health and Human Development (Bethesda, MD) from FVB/B6/129 as described previously.24 The null mutation is a lethal knockout, and the mice die shortly after birth. Therefore, each litter was removed immediately after birth and killed by chloroform inhalation and exsanguinated by decapitation after cervical dislocation. The GI tract from the proximal esophagus to

Morphological studies

Stomachs, jejunums, and colons from 81 animals were examined for the presence of enteric neurons using immunohistochemical and histochemical techniques. The pan-neuronal marker PGP 9.5 was used to determine the distribution of enteric neurons and processes. In wild-type (+/+) and heterozygous (+/−) tissues, antibodies to PGP 9.5 detected intense immunoreactivity in myenteric neurons and in interganglionic nerve bundles throughout the gastric antrum, small intestine, and colon (Figure 1A).

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Discussion

Many investigators have described electrical rhythmicity in the GI tract as non-neural in origin (for review, see Szurszewski30). This conclusion was based on the observation that treatment of the gut with tetrodotoxin, to block propagation of nerve action potentials, does not eliminate slow wave activity. The present study provides powerful support to the notion that electrical rhythmicity in the GI tract is not dependent on inputs from enteric neurons and shows that development and

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    Address requests for reprints to: Sean M. Ward, Ph.D., Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, Nevada 89557. e-mail: [email protected]; fax: (775) 784-6903.

    ☆☆

    Supported by National Institutes of Health grant DK40569. The Morphology Core Laboratory (supported by DK41315) was used for the immunohistochemical studies.

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