Limits...
Lkb1 deficiency alters goblet and paneth cell differentiation in the small intestine.

Shorning BY, Zabkiewicz J, McCarthy A, Pearson HB, Winton DJ, Sansom OJ, Ashworth A, Clarke AR - PLoS ONE (2009)

Bottom Line: Bi-allelic loss of Lkb1 led to the perturbed development of Paneth and goblet cell lineages.These changes were characterised by the lack of Delta ligand expression in Lkb1-deficient secretory cells and a significant increase in the levels of the downstream Notch signalling effector Hes5 but not Hes1.Our data show that Lkb1 is required for the normal differentiation of secretory cell lineages within the intestine, and that Lkb1 deficiency modulates Notch signalling modulation in post-mitotic cells.

View Article: PubMed Central - PubMed

Affiliation: Cardiff School of Biosciences, Cardiff University, Cardiff, United Kingdom.

ABSTRACT
The Lkb1 tumour suppressor is a multitasking kinase participating in a range of physiological processes. We have determined the impact of Lkb1 deficiency on intestinal homeostasis, particularly focussing on secretory cell differentiation and development since we observe strong expression of Lkb1 in normal small intestine Paneth and goblet cells. We crossed mice bearing an Lkb1 allele flanked with LoxP sites with those carrying a Cyp1a1-specific inducible Cre recombinase. Lkb1 was efficiently deleted from the epithelial cells of the mouse intestine after intraperitoneal injection of the inducing agent beta-naphthoflavone. Bi-allelic loss of Lkb1 led to the perturbed development of Paneth and goblet cell lineages. These changes were characterised by the lack of Delta ligand expression in Lkb1-deficient secretory cells and a significant increase in the levels of the downstream Notch signalling effector Hes5 but not Hes1. Our data show that Lkb1 is required for the normal differentiation of secretory cell lineages within the intestine, and that Lkb1 deficiency modulates Notch signalling modulation in post-mitotic cells.

Show MeSH

Related in: MedlinePlus

Lkb1-deficiency alters the expression of Notch signalling components and leads to the abnormal intestinal secretory cell differentiation.A, Western blot analysis of Notch signalling effectors, Hes1 and Hes5 at day 6 in induced Cre-negative Lkb1 fl/fl (+/+) (lanes 1, 2) and Lkb1-deficient (‘−/−’) intestinal tissue (lane 3, 4), Hes5 expression has a notable increase. B, Immunohistochemistry showing the localisation of Hes5-positive cells (pointed with red arrows) in Lkb1-deficient crypts (‘−/−’). C, Western blot analysis showing decrease in the phosphorylation of MARK kinases activation loop in Lkb1-deficient intestine (‘−/−’) while AMPK (Thr 172) phosphorylation levels are intact. D, Delta ligand (Dll1) immunostaining at day 6 in induced Cre-negative Lkb1 fl/fl (+/+) and Lkb1-deficient (‘−/−’) intestines displaying a decrease in Dll1 expression in Paneth and goblet cells of Lkb1-deficient intestines. Dll positive cells pointed with red arrows. E, Immunohistochemistry showing the localisation of Mark1 in goblet and Paneth cells both in WT (+/+) and Lkb1-deficient crypts (‘−/−’), pointed with red arrows. Scale bars correspond to 50 µm. Alcian Blue stain was performed to show mucin-secreting cells.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2626247&req=5

pone-0004264-g003: Lkb1-deficiency alters the expression of Notch signalling components and leads to the abnormal intestinal secretory cell differentiation.A, Western blot analysis of Notch signalling effectors, Hes1 and Hes5 at day 6 in induced Cre-negative Lkb1 fl/fl (+/+) (lanes 1, 2) and Lkb1-deficient (‘−/−’) intestinal tissue (lane 3, 4), Hes5 expression has a notable increase. B, Immunohistochemistry showing the localisation of Hes5-positive cells (pointed with red arrows) in Lkb1-deficient crypts (‘−/−’). C, Western blot analysis showing decrease in the phosphorylation of MARK kinases activation loop in Lkb1-deficient intestine (‘−/−’) while AMPK (Thr 172) phosphorylation levels are intact. D, Delta ligand (Dll1) immunostaining at day 6 in induced Cre-negative Lkb1 fl/fl (+/+) and Lkb1-deficient (‘−/−’) intestines displaying a decrease in Dll1 expression in Paneth and goblet cells of Lkb1-deficient intestines. Dll positive cells pointed with red arrows. E, Immunohistochemistry showing the localisation of Mark1 in goblet and Paneth cells both in WT (+/+) and Lkb1-deficient crypts (‘−/−’), pointed with red arrows. Scale bars correspond to 50 µm. Alcian Blue stain was performed to show mucin-secreting cells.

Mentions: We therefore analysed levels of the Notch signalling effectors Hes1 and Hes5 by Western blot. Hes1 levels did not significantly change (fig 3, A), but Hes5 was notably elevated (fig 3, A). Immunohistochemical analysis of control intestines at day 6 showed Hes5-positive cells to be localised predominantly at the crypt-villus junction (fig 2, B, +/+). Lkb1-deficient samples showed that Hes5 positive cells also appear at the bottom of the crypt and amongst Paneth cells (fig 2 B, ‘−/−’).


Lkb1 deficiency alters goblet and paneth cell differentiation in the small intestine.

Shorning BY, Zabkiewicz J, McCarthy A, Pearson HB, Winton DJ, Sansom OJ, Ashworth A, Clarke AR - PLoS ONE (2009)

Lkb1-deficiency alters the expression of Notch signalling components and leads to the abnormal intestinal secretory cell differentiation.A, Western blot analysis of Notch signalling effectors, Hes1 and Hes5 at day 6 in induced Cre-negative Lkb1 fl/fl (+/+) (lanes 1, 2) and Lkb1-deficient (‘−/−’) intestinal tissue (lane 3, 4), Hes5 expression has a notable increase. B, Immunohistochemistry showing the localisation of Hes5-positive cells (pointed with red arrows) in Lkb1-deficient crypts (‘−/−’). C, Western blot analysis showing decrease in the phosphorylation of MARK kinases activation loop in Lkb1-deficient intestine (‘−/−’) while AMPK (Thr 172) phosphorylation levels are intact. D, Delta ligand (Dll1) immunostaining at day 6 in induced Cre-negative Lkb1 fl/fl (+/+) and Lkb1-deficient (‘−/−’) intestines displaying a decrease in Dll1 expression in Paneth and goblet cells of Lkb1-deficient intestines. Dll positive cells pointed with red arrows. E, Immunohistochemistry showing the localisation of Mark1 in goblet and Paneth cells both in WT (+/+) and Lkb1-deficient crypts (‘−/−’), pointed with red arrows. Scale bars correspond to 50 µm. Alcian Blue stain was performed to show mucin-secreting cells.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2626247&req=5

pone-0004264-g003: Lkb1-deficiency alters the expression of Notch signalling components and leads to the abnormal intestinal secretory cell differentiation.A, Western blot analysis of Notch signalling effectors, Hes1 and Hes5 at day 6 in induced Cre-negative Lkb1 fl/fl (+/+) (lanes 1, 2) and Lkb1-deficient (‘−/−’) intestinal tissue (lane 3, 4), Hes5 expression has a notable increase. B, Immunohistochemistry showing the localisation of Hes5-positive cells (pointed with red arrows) in Lkb1-deficient crypts (‘−/−’). C, Western blot analysis showing decrease in the phosphorylation of MARK kinases activation loop in Lkb1-deficient intestine (‘−/−’) while AMPK (Thr 172) phosphorylation levels are intact. D, Delta ligand (Dll1) immunostaining at day 6 in induced Cre-negative Lkb1 fl/fl (+/+) and Lkb1-deficient (‘−/−’) intestines displaying a decrease in Dll1 expression in Paneth and goblet cells of Lkb1-deficient intestines. Dll positive cells pointed with red arrows. E, Immunohistochemistry showing the localisation of Mark1 in goblet and Paneth cells both in WT (+/+) and Lkb1-deficient crypts (‘−/−’), pointed with red arrows. Scale bars correspond to 50 µm. Alcian Blue stain was performed to show mucin-secreting cells.
Mentions: We therefore analysed levels of the Notch signalling effectors Hes1 and Hes5 by Western blot. Hes1 levels did not significantly change (fig 3, A), but Hes5 was notably elevated (fig 3, A). Immunohistochemical analysis of control intestines at day 6 showed Hes5-positive cells to be localised predominantly at the crypt-villus junction (fig 2, B, +/+). Lkb1-deficient samples showed that Hes5 positive cells also appear at the bottom of the crypt and amongst Paneth cells (fig 2 B, ‘−/−’).

Bottom Line: Bi-allelic loss of Lkb1 led to the perturbed development of Paneth and goblet cell lineages.These changes were characterised by the lack of Delta ligand expression in Lkb1-deficient secretory cells and a significant increase in the levels of the downstream Notch signalling effector Hes5 but not Hes1.Our data show that Lkb1 is required for the normal differentiation of secretory cell lineages within the intestine, and that Lkb1 deficiency modulates Notch signalling modulation in post-mitotic cells.

View Article: PubMed Central - PubMed

Affiliation: Cardiff School of Biosciences, Cardiff University, Cardiff, United Kingdom.

ABSTRACT
The Lkb1 tumour suppressor is a multitasking kinase participating in a range of physiological processes. We have determined the impact of Lkb1 deficiency on intestinal homeostasis, particularly focussing on secretory cell differentiation and development since we observe strong expression of Lkb1 in normal small intestine Paneth and goblet cells. We crossed mice bearing an Lkb1 allele flanked with LoxP sites with those carrying a Cyp1a1-specific inducible Cre recombinase. Lkb1 was efficiently deleted from the epithelial cells of the mouse intestine after intraperitoneal injection of the inducing agent beta-naphthoflavone. Bi-allelic loss of Lkb1 led to the perturbed development of Paneth and goblet cell lineages. These changes were characterised by the lack of Delta ligand expression in Lkb1-deficient secretory cells and a significant increase in the levels of the downstream Notch signalling effector Hes5 but not Hes1. Our data show that Lkb1 is required for the normal differentiation of secretory cell lineages within the intestine, and that Lkb1 deficiency modulates Notch signalling modulation in post-mitotic cells.

Show MeSH
Related in: MedlinePlus