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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

SPRY4-ir in the ICC of 3-month-old KitWT/K641E antrum.Widefield microscopy, sequential channels acquisitions. Left column: KIT immunoreactivity (-ir) ICC in WT, Spry4 KO and KitWT/K641E. Middle column: SPRY4-ir in the 3 genotypes. Right column: merged images: KIT-ir and SPRY4-ir displayed in green and in red, respectively. SPRY4-ir was detected in the KIT-ir ICC solely in KitWT/K641E mice. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm.
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pone.0124861.g001: SPRY4-ir in the ICC of 3-month-old KitWT/K641E antrum.Widefield microscopy, sequential channels acquisitions. Left column: KIT immunoreactivity (-ir) ICC in WT, Spry4 KO and KitWT/K641E. Middle column: SPRY4-ir in the 3 genotypes. Right column: merged images: KIT-ir and SPRY4-ir displayed in green and in red, respectively. SPRY4-ir was detected in the KIT-ir ICC solely in KitWT/K641E mice. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm.

Mentions: This study was based on our previous observation of elevated Spry4 expression in KitK641E/K641E antrum as compared to WT mice [19]. Here, we confirmed, in 10-day-old animals (P10), strong Spry4 expression in KitK641E/K641E antrum compared to WT littermates, while no Spry4 expression could be detected in Spry4 KO or KitK641E/K641E-Spry4 KO antrum (S1A Fig). In 3-month-old animals, SPRY4-ir was detected in the ICC of KitWT/K641E animals, but not in WT nor in Spry4 KO animals (Fig 1). These data are in line with our previous observations [19]. SPRY4 immunoreactivity (-ir) was also detected in the ICC of P10 KitK641E/K641E mice, in contrast to WT and Spry4 KO mice (S1B Fig).


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)

SPRY4-ir in the ICC of 3-month-old KitWT/K641E antrum.Widefield microscopy, sequential channels acquisitions. Left column: KIT immunoreactivity (-ir) ICC in WT, Spry4 KO and KitWT/K641E. Middle column: SPRY4-ir in the 3 genotypes. Right column: merged images: KIT-ir and SPRY4-ir displayed in green and in red, respectively. SPRY4-ir was detected in the KIT-ir ICC solely in KitWT/K641E mice. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4414615&req=5

pone.0124861.g001: SPRY4-ir in the ICC of 3-month-old KitWT/K641E antrum.Widefield microscopy, sequential channels acquisitions. Left column: KIT immunoreactivity (-ir) ICC in WT, Spry4 KO and KitWT/K641E. Middle column: SPRY4-ir in the 3 genotypes. Right column: merged images: KIT-ir and SPRY4-ir displayed in green and in red, respectively. SPRY4-ir was detected in the KIT-ir ICC solely in KitWT/K641E mice. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm.
Mentions: This study was based on our previous observation of elevated Spry4 expression in KitK641E/K641E antrum as compared to WT mice [19]. Here, we confirmed, in 10-day-old animals (P10), strong Spry4 expression in KitK641E/K641E antrum compared to WT littermates, while no Spry4 expression could be detected in Spry4 KO or KitK641E/K641E-Spry4 KO antrum (S1A Fig). In 3-month-old animals, SPRY4-ir was detected in the ICC of KitWT/K641E animals, but not in WT nor in Spry4 KO animals (Fig 1). These data are in line with our previous observations [19]. SPRY4 immunoreactivity (-ir) was also detected in the ICC of P10 KitK641E/K641E mice, in contrast to WT and Spry4 KO mice (S1B Fig).

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