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Guanylin and uroguanylin are produced by mouse intestinal epithelial cells of columnar and secretory lineage

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ABSTRACT

Guanylin (GN) and uroguanylin (UGN), through activation of guanylyl cyclase C (GCC), serve to control intestinal fluid homeostasis. Both peptides are produced in the intestinal epithelium, but their cellular origin has not been fully charted. Using quantitative PCR and an improved in situ hybridization technique (RNAscope), we have assessed the expression of GN (Guca2a), UGN (Guca2b), and GCC (Gucy2c) in mouse intestine. In the crypts of Lieberkühn, expression of Guca2a and Guca2b was restricted to cells of secretory lineage, at the crypt’s base, and to a region above, previously identified as a common origin of cellular differentiation. In this compartment, comparatively uniform levels of Guca2a and Guca2b expression were observed throughout the length of the gut. In contrast, Guca2a and Guca2b expression in the villus–surface region was more variable, and reflected the distinct, but overlapping expression pattern observed previously. Accordingly, in jejunum and ileum, Guca2a and Guca2b were abundantly expressed by enterocytes, whereas in colon only Guca2a transcript was found in the surface region. In duodenum, only low levels of Guca2b transcript were observed in columnar cells, and Guca2a expression was restricted entirely to cells of the secretory lineage. Gucy2c was shown to be expressed relatively uniformly along the rostrocaudal and crypt–villus axes and was also found in the duodenal glands. Our study reveals novel aspects of the cellular localization of the GCC signaling axis that, apart from its role in the regulation of fluid balance, link it to pH regulation, cell cycle control, and host defense.

Electronic supplementary material: The online version of this article (doi:10.1007/s00418-016-1453-4) contains supplementary material, which is available to authorized users.

No MeSH data available.


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Partitioning of Guca2a (a), Guca2b (b), and Gucy2c (c) transcripts along the rostrocaudal axis of the mouse intestinal tract. Transcript levels in 6 equidistant sections of small intestine and 2 sections of colon (see diagram) were assessed by qPCR, using expression of Gapdh as a reference. Data depict mean ± standard error. N = 6
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Fig1: Partitioning of Guca2a (a), Guca2b (b), and Gucy2c (c) transcripts along the rostrocaudal axis of the mouse intestinal tract. Transcript levels in 6 equidistant sections of small intestine and 2 sections of colon (see diagram) were assessed by qPCR, using expression of Gapdh as a reference. Data depict mean ± standard error. N = 6

Mentions: Quantitative PCR analysis showed that Guca2a transcript levels gradually increased along the rostrocaudal axis of the small intestine, and peaked in the proximal colon (Fig. 1a). In contrast, Guca2b transcript levels were low in colon. Guca2b levels were also low in the duodenum, but rose steeply along the rostrocaudal axis, and peaked in the middle to distal part of the small intestine (Fig. 1b). Gucy2c was expressed at much lower (>tenfold) levels than Guca2a or Guca2b and was partitioned more uniformly (Fig. 1c). Distribution of these transcripts was similar in male and female mice (not shown).Fig. 1


Guanylin and uroguanylin are produced by mouse intestinal epithelial cells of columnar and secretory lineage
Partitioning of Guca2a (a), Guca2b (b), and Gucy2c (c) transcripts along the rostrocaudal axis of the mouse intestinal tract. Transcript levels in 6 equidistant sections of small intestine and 2 sections of colon (see diagram) were assessed by qPCR, using expression of Gapdh as a reference. Data depict mean ± standard error. N = 6
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5037145&req=5

Fig1: Partitioning of Guca2a (a), Guca2b (b), and Gucy2c (c) transcripts along the rostrocaudal axis of the mouse intestinal tract. Transcript levels in 6 equidistant sections of small intestine and 2 sections of colon (see diagram) were assessed by qPCR, using expression of Gapdh as a reference. Data depict mean ± standard error. N = 6
Mentions: Quantitative PCR analysis showed that Guca2a transcript levels gradually increased along the rostrocaudal axis of the small intestine, and peaked in the proximal colon (Fig. 1a). In contrast, Guca2b transcript levels were low in colon. Guca2b levels were also low in the duodenum, but rose steeply along the rostrocaudal axis, and peaked in the middle to distal part of the small intestine (Fig. 1b). Gucy2c was expressed at much lower (>tenfold) levels than Guca2a or Guca2b and was partitioned more uniformly (Fig. 1c). Distribution of these transcripts was similar in male and female mice (not shown).Fig. 1

View Article: PubMed Central - PubMed

ABSTRACT

Guanylin (GN) and uroguanylin (UGN), through activation of guanylyl cyclase C (GCC), serve to control intestinal fluid homeostasis. Both peptides are produced in the intestinal epithelium, but their cellular origin has not been fully charted. Using quantitative PCR and an improved in situ hybridization technique (RNAscope), we have assessed the expression of GN (Guca2a), UGN (Guca2b), and GCC (Gucy2c) in mouse intestine. In the crypts of Lieberkühn, expression of Guca2a and Guca2b was restricted to cells of secretory lineage, at the crypt’s base, and to a region above, previously identified as a common origin of cellular differentiation. In this compartment, comparatively uniform levels of Guca2a and Guca2b expression were observed throughout the length of the gut. In contrast, Guca2a and Guca2b expression in the villus–surface region was more variable, and reflected the distinct, but overlapping expression pattern observed previously. Accordingly, in jejunum and ileum, Guca2a and Guca2b were abundantly expressed by enterocytes, whereas in colon only Guca2a transcript was found in the surface region. In duodenum, only low levels of Guca2b transcript were observed in columnar cells, and Guca2a expression was restricted entirely to cells of the secretory lineage. Gucy2c was shown to be expressed relatively uniformly along the rostrocaudal and crypt–villus axes and was also found in the duodenal glands. Our study reveals novel aspects of the cellular localization of the GCC signaling axis that, apart from its role in the regulation of fluid balance, link it to pH regulation, cell cycle control, and host defense.

Electronic supplementary material: The online version of this article (doi:10.1007/s00418-016-1453-4) contains supplementary material, which is available to authorized users.

No MeSH data available.


Related in: MedlinePlus