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Spatio-temporal expression profile of stem cell-associated gene LGR5 in the intestine during thyroid hormone-dependent metamorphosis in Xenopus laevis.

Sun G, Hasebe T, Fujimoto K, Lu R, Fu L, Matsuda H, Kajita M, Ishizuya-Oka A, Shi YB - PLoS ONE (2010)

Bottom Line: The expression of LGR5 is induced in the limb, tail, and intestine by T3 during metamorphosis.More importantly, LGR5 mRNA is localized to the developing adult epithelial stem cells of the intestine.These results suggest that LGR5-expressing cells are the stem/progenitor cells of the adult intestine and that LGR5 plays a role in the development and/or maintenance of the adult intestinal stem cells during postembryonic development in vertebrates.

View Article: PubMed Central - PubMed

Affiliation: Section on Molecular Morphogenesis, Laboratory of Gene Regulation and Development, Program in Cellular Regulation and Metabolism, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, United States of America.

ABSTRACT

Background: The intestinal epithelium undergoes constant self-renewal throughout adult life across vertebrates. This is accomplished through the proliferation and subsequent differentiation of the adult stem cells. This self-renewal system is established in the so-called postembryonic developmental period in mammals when endogenous thyroid hormone (T3) levels are high.

Methodology/principal findings: The T3-dependent metamorphosis in anurans like Xenopus laevis resembles the mammalian postembryonic development and offers a unique opportunity to study how the adult stem cells are developed. The tadpole intestine is predominantly a monolayer of larval epithelial cells. During metamorphosis, the larval epithelial cells undergo apoptosis and, concurrently, adult epithelial stem/progenitor cells develop de novo, rapidly proliferate, and then differentiate to establish a trough-crest axis of the epithelial fold, resembling the crypt-villus axis in the adult mammalian intestine. The leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5) is a well-established stem cell marker in the adult mouse intestinal crypt. Here we have cloned and analyzed the spatiotemporal expression profile of LGR5 gene during frog metamorphosis. We show that the two duplicated LGR5 genes in Xenopus laevis and the LGR5 gene in Xenopus tropicalis are highly homologous to the LGR5 in other vertebrates. The expression of LGR5 is induced in the limb, tail, and intestine by T3 during metamorphosis. More importantly, LGR5 mRNA is localized to the developing adult epithelial stem cells of the intestine.

Conclusions/significance: These results suggest that LGR5-expressing cells are the stem/progenitor cells of the adult intestine and that LGR5 plays a role in the development and/or maintenance of the adult intestinal stem cells during postembryonic development in vertebrates.

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T3-dependent expression of LGR5 mRNA in the small intestine.Cross sections of the intestine from premetamorphic tadpoles (stage 54) treated with 10 nM T3 for 1 (A, A′), 3 (B, B′) and 5 days (C, C′) were hybridized with LGR5 antisense probe. AE: adult epithelial cell including the progenitor/stem cell, CT: connective tissue, LE: larval epithelial cell, Lu: lumen, M: muscle layer. Scale bars are 100 µm (A–C) and 20 µm (A′–C′), respectively.
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pone-0013605-g006: T3-dependent expression of LGR5 mRNA in the small intestine.Cross sections of the intestine from premetamorphic tadpoles (stage 54) treated with 10 nM T3 for 1 (A, A′), 3 (B, B′) and 5 days (C, C′) were hybridized with LGR5 antisense probe. AE: adult epithelial cell including the progenitor/stem cell, CT: connective tissue, LE: larval epithelial cell, Lu: lumen, M: muscle layer. Scale bars are 100 µm (A–C) and 20 µm (A′–C′), respectively.

Mentions: Given the T3-dependence of intestinal metamorphosis and the up-regulation of LGR5 by T3, we next examined the expression patterns of LGR5 mRNA after T3 treatment of premetamorphic tadpoles at stage 54 (Fig. 6). For this experiment, tadpoles were treated with 10 nM T3 to induce precocious metamorphosis. After 1 day of T3 treatment, there was little change in the expression of LGR5 (Fig. 6A, A′). Its expression was up-regulated after 3 and 5 days of T3 treatment (Fig. 6B, B′, C, C′). The expression patterns were essentially the same as those during natural metamorphosis (compare Fig 6 with Fig. 5). That is, during both natural and T3-induced metamorphosis, the expression of LGR5 mRNA was restricted to the newly developed, proliferating adult cells but not the larval epithelial cells undergoing apoptosis, suggesting that this gene plays a role in development of the adult epithelial progenitor/stem cells and may also serve as a good molecular marker for them.


Spatio-temporal expression profile of stem cell-associated gene LGR5 in the intestine during thyroid hormone-dependent metamorphosis in Xenopus laevis.

Sun G, Hasebe T, Fujimoto K, Lu R, Fu L, Matsuda H, Kajita M, Ishizuya-Oka A, Shi YB - PLoS ONE (2010)

T3-dependent expression of LGR5 mRNA in the small intestine.Cross sections of the intestine from premetamorphic tadpoles (stage 54) treated with 10 nM T3 for 1 (A, A′), 3 (B, B′) and 5 days (C, C′) were hybridized with LGR5 antisense probe. AE: adult epithelial cell including the progenitor/stem cell, CT: connective tissue, LE: larval epithelial cell, Lu: lumen, M: muscle layer. Scale bars are 100 µm (A–C) and 20 µm (A′–C′), respectively.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2962644&req=5

pone-0013605-g006: T3-dependent expression of LGR5 mRNA in the small intestine.Cross sections of the intestine from premetamorphic tadpoles (stage 54) treated with 10 nM T3 for 1 (A, A′), 3 (B, B′) and 5 days (C, C′) were hybridized with LGR5 antisense probe. AE: adult epithelial cell including the progenitor/stem cell, CT: connective tissue, LE: larval epithelial cell, Lu: lumen, M: muscle layer. Scale bars are 100 µm (A–C) and 20 µm (A′–C′), respectively.
Mentions: Given the T3-dependence of intestinal metamorphosis and the up-regulation of LGR5 by T3, we next examined the expression patterns of LGR5 mRNA after T3 treatment of premetamorphic tadpoles at stage 54 (Fig. 6). For this experiment, tadpoles were treated with 10 nM T3 to induce precocious metamorphosis. After 1 day of T3 treatment, there was little change in the expression of LGR5 (Fig. 6A, A′). Its expression was up-regulated after 3 and 5 days of T3 treatment (Fig. 6B, B′, C, C′). The expression patterns were essentially the same as those during natural metamorphosis (compare Fig 6 with Fig. 5). That is, during both natural and T3-induced metamorphosis, the expression of LGR5 mRNA was restricted to the newly developed, proliferating adult cells but not the larval epithelial cells undergoing apoptosis, suggesting that this gene plays a role in development of the adult epithelial progenitor/stem cells and may also serve as a good molecular marker for them.

Bottom Line: The expression of LGR5 is induced in the limb, tail, and intestine by T3 during metamorphosis.More importantly, LGR5 mRNA is localized to the developing adult epithelial stem cells of the intestine.These results suggest that LGR5-expressing cells are the stem/progenitor cells of the adult intestine and that LGR5 plays a role in the development and/or maintenance of the adult intestinal stem cells during postembryonic development in vertebrates.

View Article: PubMed Central - PubMed

Affiliation: Section on Molecular Morphogenesis, Laboratory of Gene Regulation and Development, Program in Cellular Regulation and Metabolism, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland, United States of America.

ABSTRACT

Background: The intestinal epithelium undergoes constant self-renewal throughout adult life across vertebrates. This is accomplished through the proliferation and subsequent differentiation of the adult stem cells. This self-renewal system is established in the so-called postembryonic developmental period in mammals when endogenous thyroid hormone (T3) levels are high.

Methodology/principal findings: The T3-dependent metamorphosis in anurans like Xenopus laevis resembles the mammalian postembryonic development and offers a unique opportunity to study how the adult stem cells are developed. The tadpole intestine is predominantly a monolayer of larval epithelial cells. During metamorphosis, the larval epithelial cells undergo apoptosis and, concurrently, adult epithelial stem/progenitor cells develop de novo, rapidly proliferate, and then differentiate to establish a trough-crest axis of the epithelial fold, resembling the crypt-villus axis in the adult mammalian intestine. The leucine-rich repeat-containing G protein-coupled receptor 5 (LGR5) is a well-established stem cell marker in the adult mouse intestinal crypt. Here we have cloned and analyzed the spatiotemporal expression profile of LGR5 gene during frog metamorphosis. We show that the two duplicated LGR5 genes in Xenopus laevis and the LGR5 gene in Xenopus tropicalis are highly homologous to the LGR5 in other vertebrates. The expression of LGR5 is induced in the limb, tail, and intestine by T3 during metamorphosis. More importantly, LGR5 mRNA is localized to the developing adult epithelial stem cells of the intestine.

Conclusions/significance: These results suggest that LGR5-expressing cells are the stem/progenitor cells of the adult intestine and that LGR5 plays a role in the development and/or maintenance of the adult intestinal stem cells during postembryonic development in vertebrates.

Show MeSH