Clonal analysis of mouse intestinal epithelial progenitors

doi:10.1016/S0016-5085(99)70222-2

Gastroenterology

Volume 116, Issue 1, January 1999, Pages 7-14

https://doi.org/10.1016/S0016-5085(99)70222-2 Get rights and content

Abstract

Background & Aims: Little is known about the cell lineages leading from stem cells to the various terminally differentiated cell types of the intestinal epithelium. In particular, the existence and characterization of intermediate progenitor types remain open issues, which are discussed in this study. Methods: Chemical mutagenesis was used to genetically mark random intestinal epithelial cells by somatic mutation of the Dlb-1 locus. Intact epithelium was isolated at various times thereafter, and the composition, size, and location of mutant clones were scored. Results & Conclusions: Analysis of clone dynamics showed short-lived (days) progenitors (C₁, M₁, and Mix) yielding one or two cell types and long-lived (months) mucous cell progenitors (M₀), columnar cell progenitors (C₀), and pluripotential stem cells (S) capable of giving rise to all epithelial cell types. Furthermore, study of clonal dispersion, during crypt branching morphogenesis or cell migration, shows that mutant progenitors usually partition into only one of the two daughter crypts and that cells are often widely dispersed in spite of the extensive intercellular junctions in the epithelium.

GASTROENTEROLOGY 1999;116:7-14

Section snippets

Materials and methods

Twenty-three SWR and 22 F1 mice, 7-week-old males, were given an intraperitoneal injection of 250 mg/kg NEU (within 20 minutes of solvation in 10% dimethyl sulfoxide in sterile phosphate-buffered saline [PBS; Sigma Chemical Co., St. Louis, MO]) at 4 PM to 5 PM. Six SWR and 6 F1 male mice served as controls. Intact sheets of epithelium from the proximal 6–8 cm of intestine were isolated from groups of 2 mice (3 at 56 days) by ventricular perfusion with 30 mmol/L EDTA¹³ 3, 4, 7, 14, 21, 28, 42,

Results and discussion

Initially, ~3–4 of every 100 crypts contained a mutant clone, most of which were extinct by 14 days (Figure 2A). Most of these clones probably originated from short-lived progenitors including Mix, C₁, and M₁ (Figure 1). The remaining ~5 mutant clones per 1000 crypts represent long-lived progenitors of various sorts (including S, M₀, and C₀ cells). Clones observed on the villus gave a similar pattern (Figure 2B), except that a clear peak occurs at 4 days, whose ascending limb represents the

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^☆: Supported by National Science and Engineering Research Council of Canada.

^☆☆: Address requests for reprints to: Matthew Bjerknes, Department of Anatomy and Cell Biology, Medical Sciences Building, University of Toronto, Toronto, Ontario M5S-1A8, Canada. e-mail: [email protected]; fax: (416)-978-3844.

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Rapid CommunicationClonal analysis of mouse intestinal epithelial progenitors☆,☆☆

Abstract

Section snippets

Materials and methods

Results and discussion

Curr Opin Genet Dev

Mutat Res

Biophys J

Biophys J

J Theor Biol

Trends Genet

Biophys J

Cell

Biophys J

Semin Dev Biol

Pathol Res Pract

Inflammatory bowel disease and adenomas in mice expressing a dominant negative N-cadherin

Science

The chameleon within: improving antigen delivery

Science

Functional morphology of the mucosa of the small intestine

Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. V. Unitarian theory of the origin of the four epithelial cell types

Am J Anat

Rapid Communication
Clonal analysis of mouse intestinal epithelial progenitors☆,☆☆