The Ets-Domain Transcription Factor Spdef Promotes Maturation of Goblet and Paneth Cells in the Intestinal Epithelium

doi:10.1053/j.gastro.2009.06.044

Gastroenterology

Volume 137, Issue 4, October 2009, Pages 1333-1345.e3

https://doi.org/10.1053/j.gastro.2009.06.044 Get rights and content

Background & Aims

Stem cells within the intestinal epithelium generate daughter cells that undergo lineage commitment and maturation through the combined action of the Wnt and Notch signaling cascades. Both pathways, in turn, regulate transcription factor networks that further define differentiation toward either enterocytes or 1 of 3 secretory cell lineages (Paneth, goblet, or enteroendocrine cells). In this study, we investigated the role of the Wnt-responsive, Ets-domain transcription factor Spdef in the differentiation of goblet and Paneth cells.

Methods

The in vivo function of Spdef was examined by disrupting the Spdef gene in mice (Spdef^−/− mice) and analyzing the intestinal phenotype using a range of histologic techniques and DNA microarray profiling.

Results

In accordance with expression data, we found that loss of Spdef severely impaired the maturation of goblet and Paneth cells and, conversely, led to an accumulation of immature secretory progenitors. Spdef appears to positively and negatively regulate a specific subset of goblet and Paneth cell genes, including Cryptdins, Mmp7, Ang4, Kallikreins, and Muc2.

Conclusions

Spdef acts downstream of Math1 to promote terminal differentiation of a secretory progenitor pool into Paneth and goblet cells.

Section snippets

Spdef Targeting Construct and Generation of Spdef^−/− Mice

The Spdef targeting construct was generated using a recombineering strategy initially described elsewhere (http://recombineering.ncifcrf.gov). Briefly, starting from BAC clone, a 12-kilobase (kb) fragment of genomic DNA comprising exon 6 of the Spdef gene was retrieved in the pl253 plasmid using the SW102 recombinogenic strain. Similarly, loxP sites flanking exon 6 and a neo-cassette were recombined into the retrieved 12-kb genomic fragment using the pl451 plasmid as a template. The targeting

Identification of Spdef as a Marker of Gut Progenitors and Mature Goblet and Paneth Cells

To shed light on the downstream gene targets regulated by Wnt signals in the gut intestine, we previously performed microarray profiling on fetal small intestines derived from wild-type vs Tcf4^−/− embryos.¹⁰ One of the target genes that attracted our attention was the transcription factor Spdef (also termed PDEF). By in situ hybridization, we found that, at late fetal stages when the gut epithelium begins to differentiate, Spdef messenger RNA (mRNA) was localized in a limited number of cells

Discussion

In this study, we showed that the Ets domain transcription factor Spdef is expressed in cells of the goblet and Paneth cell lineages, as well as in early crypt progenitor cells. In numerous crypts, we also detected Spdef at or around the +4 position just above the Paneth cell compartment, suggesting that these cells may represent immediate descendants of either Bmi1⁺ and/or Lgr5⁺ stem cells.24, 25 In support of this claim, we have recently found through microarray profiling that Spdef is

Acknowledgments

The authors thank Dr Jeffrey Whitsett for generously providing us with the Spdef antibody and Stieneke van den Brink for excellent technical assistance with ES cell experiments.

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: A.G.'s present address is Samuel Lunenfeld Research Institute, 1070-600 University Avenue, M5G 1X5 Toronto, Ontario, Canada.

: Conflicts of interest The authors disclose no conflicts.

: Funding Supported by Koninklijke Nederlandse Akademie van Wetenschappen (KNAW) and Koningin Wilhelmina Fonds (KWF).

View full text

Basic—Alimentary TractThe Ets-Domain Transcription Factor Spdef Promotes Maturation of Goblet and Paneth Cells in the Intestinal Epithelium

Background & Aims

Methods

Results

Conclusions

Section snippets

Spdef Targeting Construct and Generation of Spdef−/− Mice

Identification of Spdef as a Marker of Gut Progenitors and Mature Goblet and Paneth Cells

Discussion

Acknowledgments

Gastroenterology

J Biol Chem

Gastroenterology

J Biol Chem

Differentiation

Cell

Stem cells, self-renewal, and differentiation in the intestinal epithelium

Annu Rev Physiol

Canonical wnt signals are essential for homeostasis of the intestinal epithelium

Genes Dev

Notch/γ-secretase inhibition turns proliferative cells in intestinal crypts and adenomas into goblet cells

Nature

Notch signals control the fate of immature progenitor cells in the intestine

Nature

Notch signaling regulates the differentiation of post-mitotic intestinal epithelial cells

Genes Dev

Direct regulation of intestinal fate by notch

Proc Natl Acad Sci U S A

Control of endodermal endocrine development by Hes-1

Nat Genet

Requirement of Math1 for secretory cell lineage commitment in the mouse intestine

Science

Wnt signalling induces maturation of Paneth cells in intestinal crypts

Nat Cell Biol

Basic—Alimentary Tract
The Ets-Domain Transcription Factor Spdef Promotes Maturation of Goblet and Paneth Cells in the Intestinal Epithelium

Spdef Targeting Construct and Generation of Spdef^−/− Mice