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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

ICC hyperplasia in the antrum of 3-month-old Spry4 KO and KitWT/K641E.A) Widefield microscopy acquisitions. PDE3A immunoreactivity (-ir) highlights ICC in the antrum of 3-month-old WT, Spry4 KO and KitWT/K641E mice. B) Ratio of PDE3A-ir ICC area in the antrum muscularis propria. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm. P-values (Kruskal-Wallis with Dunn’s post hoc) *: p<0.05, **: p<0.01
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pone.0124861.g002: ICC hyperplasia in the antrum of 3-month-old Spry4 KO and KitWT/K641E.A) Widefield microscopy acquisitions. PDE3A immunoreactivity (-ir) highlights ICC in the antrum of 3-month-old WT, Spry4 KO and KitWT/K641E mice. B) Ratio of PDE3A-ir ICC area in the antrum muscularis propria. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm. P-values (Kruskal-Wallis with Dunn’s post hoc) *: p<0.05, **: p<0.01

Mentions: Quantification of the PDE3A-ir ICC density in the musculature was performed on the entire circumference of the antrum, by normalizing the area of PDE3A-ir positive signal to the total muscularis propria area. Compared to WT littermates, 3-month-old Spry4 KO animals showed a significant increase in PDE3A-ir ICC area, similar to KitWT/K641E mice (Fig 2) while in P10 Spry4 KO animals, no difference was observed in the PDE3A-ir ICC area compared to their WT littermates (S5 Fig). In contrast, P10 homozygous KitK641E/K641E already exhibited a massive PDE3A-ir ICC hyperplasia (S5 Fig), replacing the entire longitudinal muscle layer in the antrum. As these homozygous mice died before weaning, heterozygous KitWT/K641E mice, which display a milder phenotype of ICC hyperplasia, were used for subsequent comparative studies in adult animals.


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)

ICC hyperplasia in the antrum of 3-month-old Spry4 KO and KitWT/K641E.A) Widefield microscopy acquisitions. PDE3A immunoreactivity (-ir) highlights ICC in the antrum of 3-month-old WT, Spry4 KO and KitWT/K641E mice. B) Ratio of PDE3A-ir ICC area in the antrum muscularis propria. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm. P-values (Kruskal-Wallis with Dunn’s post hoc) *: p<0.05, **: p<0.01
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4414615&req=5

pone.0124861.g002: ICC hyperplasia in the antrum of 3-month-old Spry4 KO and KitWT/K641E.A) Widefield microscopy acquisitions. PDE3A immunoreactivity (-ir) highlights ICC in the antrum of 3-month-old WT, Spry4 KO and KitWT/K641E mice. B) Ratio of PDE3A-ir ICC area in the antrum muscularis propria. Abbreviations: LM: longitudinal muscle layer, CM: circular muscle layer, *: location of myenteric plexus, scale bar: 100μm. P-values (Kruskal-Wallis with Dunn’s post hoc) *: p<0.05, **: p<0.01
Mentions: Quantification of the PDE3A-ir ICC density in the musculature was performed on the entire circumference of the antrum, by normalizing the area of PDE3A-ir positive signal to the total muscularis propria area. Compared to WT littermates, 3-month-old Spry4 KO animals showed a significant increase in PDE3A-ir ICC area, similar to KitWT/K641E mice (Fig 2) while in P10 Spry4 KO animals, no difference was observed in the PDE3A-ir ICC area compared to their WT littermates (S5 Fig). In contrast, P10 homozygous KitK641E/K641E already exhibited a massive PDE3A-ir ICC hyperplasia (S5 Fig), replacing the entire longitudinal muscle layer in the antrum. As these homozygous mice died before weaning, heterozygous KitWT/K641E mice, which display a milder phenotype of ICC hyperplasia, were used for subsequent comparative studies in adult animals.

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