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Hyperplasia of interstitial cells of cajal in sprouty homolog 4 deficient mice.

Thys A, Vandenberghe P, Hague P, Klein OD, Erneux C, Vanderwinden JM - PLoS ONE (2015)

Bottom Line: Sprouty homolog 4 was upregulated both at the mRNA and protein level in these cells, suggesting that Sprouty homolog 4 is downstream of oncogenic KIT activation and potentially engaged in the negative feedback loop of ERK activation in this model.Here, we used KitK641E heterozygous and Sprouty homolog 4 knock out animals to quantify interstitial cells of Cajal in situ, using quantitative immunofluorescence for the receptor tyrosine kinase Kit and for phosphodiesterase 3a (PDE3A).We concluded that the lack of Sprouty homolog 4 expression leads to hyperplasia of the interstitial cells of Cajal and is functionally associated with a delayed transit time.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Neurophysiology, Faculty of Medicine, Université Libre de Bruxelles, Brussels, Belgium.

ABSTRACT
Gastrointestinal stromal tumors, which are thought to derive from interstitial cells of Cajal or their precursors, often harbor an oncogenic mutation of the KIT receptor tyrosine kinase. Sprouty homolog 4, a known negative regulator of ERK pathway, has been identified in the interstitial cells of Cajal in the KitK641E murine model of gastrointestinal stromal tumors. Sprouty homolog 4 was upregulated both at the mRNA and protein level in these cells, suggesting that Sprouty homolog 4 is downstream of oncogenic KIT activation and potentially engaged in the negative feedback loop of ERK activation in this model. Here, we used KitK641E heterozygous and Sprouty homolog 4 knock out animals to quantify interstitial cells of Cajal in situ, using quantitative immunofluorescence for the receptor tyrosine kinase Kit and for phosphodiesterase 3a (PDE3A). In the antrum of Sprouty homolog 4 knock out mice, hyperplasia of interstitial cells of Cajal was reminiscent of the KitK641E heterozygous mice antrum. Additionally, the density of interstitial cells of Cajal was higher in the colon of adult Sprouty homolog 4 knock out mice than in WT littermates, although hyperplasia seemed more severe in KitK641E heterozygous mice. Functional transit studies also show similarities between Sprouty homolog 4 knock out and KitK641E heterozygous mice, as the total transit time in 9 month old animals was significantly increased in both genotypes compared to WT littermates. We concluded that the lack of Sprouty homolog 4 expression leads to hyperplasia of the interstitial cells of Cajal and is functionally associated with a delayed transit time.

No MeSH data available.


Related in: MedlinePlus

No detectable pp70S6 in ICC of antrum of WT, Spry4 KO or KitWT/K641E.Widefield microscopy, sequential channels acquisitions. Left column: PDE3A immunoreactivity (-ir) ICC in WT, Spry4 KO and KitWT/K641E antrum. Middle column: pp70S6-ir for each genotype. Right column: merged images: PDE3A and pp70S6-ir displayed in green and red, respectively. pp70S6 was consistently detected in myenteric plexus and nerve fibers in the muscularis propria but not in PDE3A-ir ICC. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm
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pone.0124861.g006: No detectable pp70S6 in ICC of antrum of WT, Spry4 KO or KitWT/K641E.Widefield microscopy, sequential channels acquisitions. Left column: PDE3A immunoreactivity (-ir) ICC in WT, Spry4 KO and KitWT/K641E antrum. Middle column: pp70S6-ir for each genotype. Right column: merged images: PDE3A and pp70S6-ir displayed in green and red, respectively. pp70S6 was consistently detected in myenteric plexus and nerve fibers in the muscularis propria but not in PDE3A-ir ICC. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm

Mentions: To further investigate a possible involvement of mTOR pathway, we performed IF for the active form of p70S6, a protein downstream of mTOR, phospho-p70S6 (pp70S6). pp70S6-ir. Was detected in the hyperplastic PDE3A-ir ICC layer of P10 KitK641E/K641E animals (S9 Fig) but, in all other genotypes, at both P10 and 3 month old, pp70S6-ir was detected only in the myenteric plexus of the antrum but not in PDE3A-ir ICC (Fig 6).


Hyperplasia of interstitial cells of cajal in sprouty homolog 4 deficient mice.

Thys A, Vandenberghe P, Hague P, Klein OD, Erneux C, Vanderwinden JM - PLoS ONE (2015)

No detectable pp70S6 in ICC of antrum of WT, Spry4 KO or KitWT/K641E.Widefield microscopy, sequential channels acquisitions. Left column: PDE3A immunoreactivity (-ir) ICC in WT, Spry4 KO and KitWT/K641E antrum. Middle column: pp70S6-ir for each genotype. Right column: merged images: PDE3A and pp70S6-ir displayed in green and red, respectively. pp70S6 was consistently detected in myenteric plexus and nerve fibers in the muscularis propria but not in PDE3A-ir ICC. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4414615&req=5

pone.0124861.g006: No detectable pp70S6 in ICC of antrum of WT, Spry4 KO or KitWT/K641E.Widefield microscopy, sequential channels acquisitions. Left column: PDE3A immunoreactivity (-ir) ICC in WT, Spry4 KO and KitWT/K641E antrum. Middle column: pp70S6-ir for each genotype. Right column: merged images: PDE3A and pp70S6-ir displayed in green and red, respectively. pp70S6 was consistently detected in myenteric plexus and nerve fibers in the muscularis propria but not in PDE3A-ir ICC. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm
Mentions: To further investigate a possible involvement of mTOR pathway, we performed IF for the active form of p70S6, a protein downstream of mTOR, phospho-p70S6 (pp70S6). pp70S6-ir. Was detected in the hyperplastic PDE3A-ir ICC layer of P10 KitK641E/K641E animals (S9 Fig) but, in all other genotypes, at both P10 and 3 month old, pp70S6-ir was detected only in the myenteric plexus of the antrum but not in PDE3A-ir ICC (Fig 6).

Bottom Line: Sprouty homolog 4 was upregulated both at the mRNA and protein level in these cells, suggesting that Sprouty homolog 4 is downstream of oncogenic KIT activation and potentially engaged in the negative feedback loop of ERK activation in this model.Here, we used KitK641E heterozygous and Sprouty homolog 4 knock out animals to quantify interstitial cells of Cajal in situ, using quantitative immunofluorescence for the receptor tyrosine kinase Kit and for phosphodiesterase 3a (PDE3A).We concluded that the lack of Sprouty homolog 4 expression leads to hyperplasia of the interstitial cells of Cajal and is functionally associated with a delayed transit time.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Neurophysiology, Faculty of Medicine, Université Libre de Bruxelles, Brussels, Belgium.

ABSTRACT
Gastrointestinal stromal tumors, which are thought to derive from interstitial cells of Cajal or their precursors, often harbor an oncogenic mutation of the KIT receptor tyrosine kinase. Sprouty homolog 4, a known negative regulator of ERK pathway, has been identified in the interstitial cells of Cajal in the KitK641E murine model of gastrointestinal stromal tumors. Sprouty homolog 4 was upregulated both at the mRNA and protein level in these cells, suggesting that Sprouty homolog 4 is downstream of oncogenic KIT activation and potentially engaged in the negative feedback loop of ERK activation in this model. Here, we used KitK641E heterozygous and Sprouty homolog 4 knock out animals to quantify interstitial cells of Cajal in situ, using quantitative immunofluorescence for the receptor tyrosine kinase Kit and for phosphodiesterase 3a (PDE3A). In the antrum of Sprouty homolog 4 knock out mice, hyperplasia of interstitial cells of Cajal was reminiscent of the KitK641E heterozygous mice antrum. Additionally, the density of interstitial cells of Cajal was higher in the colon of adult Sprouty homolog 4 knock out mice than in WT littermates, although hyperplasia seemed more severe in KitK641E heterozygous mice. Functional transit studies also show similarities between Sprouty homolog 4 knock out and KitK641E heterozygous mice, as the total transit time in 9 month old animals was significantly increased in both genotypes compared to WT littermates. We concluded that the lack of Sprouty homolog 4 expression leads to hyperplasia of the interstitial cells of Cajal and is functionally associated with a delayed transit time.

No MeSH data available.


Related in: MedlinePlus