Wnt-reporter expression pattern in the mouse intestine during homeostasis

Paige Davies, Adria D. Dismuke, Anne E. Powell, Kevin H. Carroll, Melissa Wong

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Background: The canonical Wnt signaling pathway is a known regulator of cell proliferation during development and maintenance of the intestinal epithelium. Perturbations in this pathway lead to aberrant epithelial proliferation and intestinal cancer. In the mature intestine, proliferation is confined to the relatively quiescent stem cells and the rapidly cycling transient-amplifying cells in the intestinal crypts. Although the Wnt signal is believed to regulate all proliferating intestinal cells, surprisingly, this has not been thoroughly demonstrated. This important determination has implications on intestinal function, especially during epithelial expansion and regeneration, and warrants an extensive characterization of Wnt-activated cells. Methods: To identify intestinal epithelial cells that actively receive a Wnt signal, we analyzed intestinal Wnt-reporter expression patterns in two different mouse lines using immunohistochemistry, enzymatic activity, in situ hybridization and qRT-PCR, then corroborated results with reporter-independent analyses. Wnt-receiving cells were further characterized for co-expression of proliferation markers, putative stem cell markers and cellular differentiation markers using an immunohistochemical approach. Finally, to demonstrate that Wnt-reporter mice have utility in detecting perturbations in intestinal Wnt signaling, the reporter response to gamma-irradiation was examined. Results: Wnt-activated cells were primarily restricted to the base of the small intestinal and colonic crypts, and were highest in numbers in the proximal small intestine, decreasing in frequency in a gradient toward the large intestine. Interestingly, the majority of the Wnt-reporter-expressing cells did not overlap with the transient-amplifying cell population. Further, while Wnt-activated cells expressed the putative stem cell marker Musashi-1, they did not co-express DCAMKL-1 or cell differentiation markers. Finally, gamma-irradiation stimulated an increase in Wnt-activated intestinal crypt cells. Conclusion: We show, for the first time, detailed characterization of the intestine from Wnt-reporter mice. Further, our data show that the majority of Wnt-receiving cells reside in the stem cell niche of the crypt base and do not extend into the proliferative transient-amplifying cell population. We also show that the Wnt-reporter mice can be used to detect changes in intestinal epithelial Wnt signaling upon physiologic injury. Our findings have an important impact on understanding the regulation of the intestinal stem cell hierarchy during homeostasis and in disease states.

Original languageEnglish (US)
Article number57
JournalBMC Gastroenterology
Volume8
DOIs
StatePublished - Dec 2 2008

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Intestines
Homeostasis
Stem Cells
Wnt Signaling Pathway
Differentiation Antigens
Stem Cell Niche
Intestinal Neoplasms
Large Intestine
Intestinal Mucosa
Population
Small Intestine
In Situ Hybridization
Regeneration
Cell Differentiation
Epithelial Cells
Immunohistochemistry
Maintenance
Cell Proliferation
Polymerase Chain Reaction
Wounds and Injuries

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Wnt-reporter expression pattern in the mouse intestine during homeostasis. / Davies, Paige; Dismuke, Adria D.; Powell, Anne E.; Carroll, Kevin H.; Wong, Melissa.

In: BMC Gastroenterology, Vol. 8, 57, 02.12.2008.

Research output: Contribution to journalArticle

Davies, Paige ; Dismuke, Adria D. ; Powell, Anne E. ; Carroll, Kevin H. ; Wong, Melissa. / Wnt-reporter expression pattern in the mouse intestine during homeostasis. In: BMC Gastroenterology. 2008 ; Vol. 8.
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